CRISPR Plasmids: Bacteria

The following CRISPR plasmids have been designed for use in bacteria.

Cut

Fully functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism which frequently causes insertions or deletions (indels) in the DNA. Indels often lead to frameshifts, creating loss of function alleles.

To introduce specific genomic changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the cell may repair a DSB using homology-directed repair (HDR) instead of NHEJ. In most experimental systems, HDR occurs at a much lower efficiency than NHEJ.

	Plasmid	Gene/Insert	Promoter	PI	Publication	Hidden Extra Search Info
	pMJ806	Cas9	T7	Doudna	A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012 Jun 28.	pEC-K-MBP
	pCas9	tracr/Cas9		Marraffini	RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2508.	Bacterial expression of Cas9 nuclease, tracrRNA and crRNA guide pACYC184
	pwtCas9-bacteria	wild-type Cas9	pLtetO-1	Qi	Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022.	aTc-inducible expression of wild-type Cas9 (S .pyogenes) for bacterial gene knockdown pUC19
	pET-28b-Cas9-His	Cas9 (Other)		Schier	Efficient mutagenesis by Cas9 protein-mediated oligonucleotide insertion and large-scale assessment of single-guide RNAs. PLoS One. 2014 May 29;9(5):e98186. doi: 10.1371/journal.pone.0098186. eCollection 2014.	For in vitro expression and purification of Cas9 protein pET-28b
	DS-SPcas	Cas9 (Other), tracrRNA precursor (Other)	proC, tracrRNA promoter	Church	Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.	Bacterial S. pyogenes Cas9 (SP) + tracrRNA expression, cloDF13/spectinomycin cloDF13-aadA
	DS-NMcas	Cas9 (Other), NM tracrRNA (Other)	proC, NM tracrRNA promoter	Church	Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.	Bacterial N. meningitidis Cas9 (NM) + tracrRNA expression, cloDF13/spectinomycin cloDF13-aadA
	DS-ST1cas	Cas9 (Other), ST1 tracrRNA (Other)	proC	Church	Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.	Bacterial S. thermophilus #1 Cas9 (ST1) + tracrRNA expression, cloDF13/spectinomycin cloDF13-aadA
	pET28a/Cas9-Cys	Cas9-Cys (Other)	T7 Promoter	Kim	Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA. Genome Res. 2014 Apr 2.	Expresses N-terminal His tag fused human codon-optimized Cas9 nuclease having C-terminal Cysteine pET28a
	pCRISPomyces-1	sSpCas9 (Synthetic), tracrRNA (Other), crRNA cassette (Synthetic)	rpsLp(XC)-BbsI, rpsLp(CF), gapdhp(EL)	Zhao	High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System. ACS Synth Biol. 2014 Dec 8.	Streptomyces expression of codon-optimized Cas9, tracrRNA, and custom crRNA pKC1139
	pCRISPomyces-2	sSpCas9 (Synthetic), gRNA cassette (Synthetic)	rpsL(XC)-BbsI, gapdhp(EL)	Zhao	High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System. ACS Synth Biol. 2014 Dec 8.	Streptomyces expression of codon-optimized Cas9 and custom gRNA pKC1139
	pCas	cas9 (Other)	native cas9 promoter	Yang	Multigene editing in the Escherichia coli genome using the CRISPR-Cas9 system. Appl Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14.	Constitutive expression of cas9 and inducible expression of lambda RED and sgR pKD46
	pET-(-30)dGFP-9xGGS-Cath-NLS-Cas9-6xHis	(-30)dGFP-9xGGS-Cath-NLS-Cas9-6xHis (Other)	T7	Liu	Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30.	Expression of (-30)dGFP-9xGGS-Cath-NLS-Cas9-6xHis in bacterial cells pET29
	pET-Cas9-6xHis	Cas9_6xHis (Other)	T7	Liu	Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30.	Expression of Cas9-6xHis in bacterial cells pET29
	pTrex-b-NLS-hSpCas9	NLS-hSpCas9	Ribo-HX1	Tarleton	CRISPR-Cas9-Mediated Single-Gene and Gene Family Disruption in Trypanosoma cruzi. MBio. 2014 Dec 30;6(1). pii: e02097-14. doi: 10.1128/mBio.02097-14.	Expression of Cas9 in T. cruzi pTrex-b
	pKCcas9dO	Scocas9 (Other), sgRNA (), act-orf4 homology arm	ptipA, j23119	Jiang	One-step high-efficiency CRISPR/Cas9-mediated genome editing in Streptomyces. Acta Biochim Biophys Sin (Shanghai). 2015 Apr;47(4):231-43. doi: 10.1093/abbs/gmv007. Epub 2015 Mar 3.	High efficiency Streptomyces genome editing by CRISPR/Cas9 system pKC1139
	pCas9-CR4	Cas9 nuclease		Prather	The no-SCAR (Scarless Cas9 Assisted Recombineering) system for genome editing in Escherichia coli. Sci Rep. 2015 Oct 14;5:15096. doi: 10.1038/srep15096.	Cas9 nuclease under control of pTet promoter with ssrA tag and constitutive tetR pdCas9-bacteria
	SP-Cas9	SP-CAS9 (Other)	T7	Geijsen	Efficient Intracellular Delivery of Native Proteins. Cell. 2015 Apr 23;161(3):674-690. doi: 10.1016/j.cell.2015.03.028.	Expresses SP-Cas9 protein in bacterial cells pET-15
	pCMK	Cas9 (Synthetic)		Rodrigue	Bacterial constitutive gRNA expression plasmid (unpublished)	Bacterial constitutive S. pyogenes Cas9 expression plasmid pCMK
	pET-Cas9-NLS-6xHis	Cas9-NLS-6xHis (Other)	T7	Liu	Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30.	Expression of Cas9-NLS-6xHis in bacterial cells pET29
	pET-NLS-Cas9-6xHis	NLS-Cas9-6xHis (Other)	T7	Liu	Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30.	Expression of NLS-Cas9-6xHis in bacterial cells pET29
	pLPhygCAS9	Humanized Streptococcus pyogenes Cas9 from PX330 (Addgene) (Other)		Matlashewski	CRISPR-Cas9-Mediated Genome Editing in Leishmania donovani. MBio. 2015 Jul 21;6(4). pii: e00861-15. doi: 10.1128/mBio.00861-15.	Expresses CAS9 in Leishmania pLPHyg2
	pYTK036	Cas9 (Other)		Dueber	A Highly-characterized Yeast Toolkit for Modular, Multi-part Assembly. ACS Synth Biol. 2015 Apr 14.	Encodes Cas9 as a Type 3 part to be used in the Dueber YTK system pYTK001
	BPK764	mammalian codon-optimized Streptococcus pyogenes Cas9-NLS-3XFlag, and SpCas9 gRNA (Other)	T7 (x2)	Joung	Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592.	Bacterial expression plasmid for SpCas9 & sgRNA (need to clone spacer into BsaI sites): T7-humanSpCas9-NLS-3xFLAG-T7-BsaIcassette-Sp-sgRNA pACYCDuet-1 Tags / Fusion Proteins NLS (C terminal on insert) 3x FLAG (C terminal on insert)
	MSP1673	mammalian codon-optimized Streptococcus thermophilus1 Cas9-NLS, and St1Cas9 gRNA (Other)	T7 (x2)	Joung	Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592.	Bacterial expression plasmid for S. thermophilus1 Cas9 & sgRNA (need to clone in spacer into BspMI sites): T7-humanSt1Cas9-NLS-T7-BspMIcassette-St1-sgRNA pACYCDuet-1 Tag / Fusion Protein NLS (C terminal on insert)
	BPK2101	mammalian codon-optimized Staphylococcus aureus Cas9-NLS-3xFlag, and SaCas9 gRNA (Other)	T7 (x2)	Joung	Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592.	Bacterial expression plasmid for S. aureus Cas9 & sgRNA (need to clone in spacer into BsaI sites): T7-humanSaCas9-NLS-3xFLAG-T7-BsaIcassette-Sa-sgRNA pACYCDuet-1 Tags / Fusion Proteins NLS (C terminal on insert) 3x FLAG (C terminal on insert)
	pHO4d-Cas9	Cas9 (Other)	T7	Nonet	Landscape of target:guide homology effects on Cas9-mediated cleavage. Nucleic Acids Res. 2014 Dec 16;42(22):13778-87. doi: 10.1093/nar/gku1102. Epub 2014 Nov 15.	Cas9nlsHis6 Bacterial Expression construct pHO4D
	pMJ915	cas9 (Other)	T7	Doudna	Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery. Elife. 2014 Dec 15;3:e04766. doi: 10.7554/eLife.04766.	Express Streptococcus pyogenes Cas9 carrying two C-terminal SV40 NLS pML-2CT
	MSP2283	mammalian codon-optimized Staphylococcus aureus Cas9-NLS-3xFlag (Other), SaCas9 sgRNA (+84)	T7, T7	Joung	Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404.	Bacterial expression plasmid for SaCas9 & sgRNA targeted to site 1: T7-humanSaCas9-NLS-3xFLAG-T7-Sa-sgRNA(84) #1 pACYCDuet-1
	MSP2253	mammalian codon-optimized KKH variant Staphylococcus aureus Cas9(E782K/N968K/R1015H)-NLS-3xFlag (Other), SaCas9 sgRNA (+84)	T7, T7	Joung	Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404.	Bacterial expression plasmid for KKH SaCas9 & sgRNA targeted to site 1: T7-humanSaCas9(E782K/N968K/R1015H)-NLS-3xFLAG-T7-Sa-sgRNA(84) #1 pACYCDuet-1
	MSP2266	mammalian codon-optimized Staphylococcus aureus Cas9-NLS-3xFlag (Other), SaCas9 sgRNA (+84)	T7, T7	Joung	Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404.	Bacterial expression plasmid for SaCas9 & sgRNA targeted to site 2: T7-humanSaCas9-NLS-3xFLAG-T7-Sa-sgRNA(84) #2 pACYCDuet-1
	MSP2292	mammalian codon-optimized KKH variant Staphylococcus aureus Cas9(E782K/N968K/R1015H)-NLS-3xFlag, and SaCas9 sgRNA(84) (Other), SaCas9 sgRNA (+84)	T7, T7	Joung	Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404.	Bacterial expression plasmid for KKH SaCas9 & sgRNA targeted to site 2: T7-humanSaCas9(E782K/N968K/R1015H)-NLS-3xFLAG-T7-Sa-sgRNA(84) #2 pACYCDuet-1
	pCas9_sgRNA_0	U6 promoter (Other), cas9 (Synthetic), tnos terminator (Other)	U6 from Ustilago maydis, synthetic otef promoter (Spellig et al., 1996, Mol. Gen. Genet. 252)	Kahmann	Genome editing in Ustilago maydis using the CRISPR-Cas system. Fungal Genet Biol. 2015 Sep 11. pii: S1087-1845(15)30025-6. doi: 10.1016/j.fgb.2015.09.001.	expresses Ustilago maydis codon-optimized Cas9, contains U. maydis U6 promoter, is self-replicating pNEBUC
	pMCSG7-Wt-NmeCas9	pMCSG7-WT-NmeCas9 (Other)		Sontheimer	DNase H Activity of Neisseria meningitidis Cas9. Mol Cell. 2015 Oct 15;60(2):242-55. doi: 10.1016/j.molcel.2015.09.020.	bacterial pMCSG7 expression vector expressing Wildtype Nme cas9 with T7 promoter, N-terminal His tag and TEV site pMCSG7
	pCas9cur	Cas9 (Other), Plasmid curing system	pBAD	Chen	Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing. Metab Eng. 2015 Sep;31:13-21. doi: 10.1016/j.ymben.2015.06.006. Epub 2015 Jun 30.	Constitutive expression of Cas9 gene and inducible expression of the plasmid curing system for CRISPR mediated genome editing of E. coli. pSC101ts
	pREDCas9	Cas9 (Other), lambda red genes, plasmid curing system	pBAD	Chen	Metabolic engineering of Escherichia coli using CRISPR-Cas9 meditated genome editing. Metab Eng. 2015 Sep;31:13-21. doi: 10.1016/j.ymben.2015.06.006. Epub 2015 Jun 30.	Constitutive expression of Cas9, inducible expression of the Red recombineering system, and inducible expression of the plasmid curing system for CRISPR mediated genome editing of E. coli. pSC101ts
	pCAS1yl	Cas9 (Other), sgRNA (Synthetic)	unknown	Yang	Multiplex gene editing of the Yarrowia lipolytica genome using the CRISPR-Cas9 system. J Ind Microbiol Biotechnol. 2016 Aug;43(8):1085-93. doi: 10.1007/s10295-016-1789-8. Epub 2016 Jun 27.	Constitutive expression of Cas9 and sgRNA in Yarrowia lipolytica cells pMCSCen1
	pMA7CR_2.0	cas9 (Other), lambda beta (Other), dam (Other), recX (Other)	pTet, pAra, pAra, pTet	Nielsen	CRMAGE: CRISPR Optimized MAGE Recombineering. Sci Rep. 2016 Jan 22;6:19452. doi: 10.1038/srep19452.	pMA7CR_2.0 contains arabinose inducible λ/RED β-protein and dam, and aTc inducible CRISPR/Cas9 and recX pBAD24
	pDEST-hisMBP-AsCpf1-EC	AsCpf1 (Synthetic)	tac promoter	Kim	Targeted mutagenesis in mice by electroporation of Cpf1 ribonucleoproteins. Nat Biotechnol. 2016 Jun 6. doi: 10.1038/nbt.3596.	Expressing his-MBP tagged AsCpf1 (E.coli codon optimized) pDEST-hisMBP
	pMAL-his-LbCpf1-EC	LbCpf1 (Synthetic)	tac promoter	Kim	Genome-wide analysis reveals specificities of Cpf1 endonucleases in human cells. Nat Biotechnol. 2016 Jun 6. doi: 10.1038/nbt.3609.	Bacterial expression - MBP-his tagged LbCpf1 (E.coli codon optimized) pMAL-c5X
	pCas9-CAT	Cas9 (Other), Chloramphenicol acetyltransferase	TgTUB1, TgTUB1	Lourido	A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 10.1016/j.cell.2016.08.019. Epub 2016 Sep 2.	Encodes Cas9 and chloramphenicol acetyltransferase (CAT) pU6-Universal
	pU6-Decoy	Decoy sgRNA	U6	Lourido	A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 10.1016/j.cell.2016.08.019. Epub 2016 Sep 2.	Encodes Cas9 and a CRISPR sgRNA that alleviates toxicity to Cas9 in Toxoplasma gondii pU6-Universal
	bbCas9pluspAAA	Cas9 (Other)	T7	Huijbers	Direct Generation of Conditional Alleles Using CRISPR/Cas9 in Mouse Zygotes. Methods Mol Biol. 2017;1642:21-35. doi: 10.1007/978-1-4939-7169-5_2.	The plasmid encodes the T7 promoter, Cas9 (from pX330 #42230) and a stretch of poly-A. After linearization with restriction enzyme SapI It is used to produce in vitro transcribed mRNA pUC
	pEC-K-CBP_CjeCas9	CjeCas9 (Other)		Doudna	Single-Stranded DNA Cleavage by Divergent CRISPR-Cas9 Enzymes. Mol Cell. 2015 Nov 5;60(3):398-407. doi: 10.1016/j.molcel.2015.10.030.	Expresses Campylobacter jejuni Cas9 (Cje Cas9). pEC-K
	p2CT-Cdi	CdiCas9		Doudna	Single-Stranded DNA Cleavage by Divergent CRISPR-Cas9 Enzymes. Mol Cell. 2015 Nov 5;60(3):398-407. doi: 10.1016/j.molcel.2015.10.030.	Expresses Corynebacterium diphtheriae Cas9 (CdiCas9) PET
	pLdCN	gRNA and Cas9 (Other)	L. donovani ribosome RNA promoter	Matlashewski	Optimized CRISPR-Cas9 Genome Editing for Leishmania and Its Use To Target a Multigene Family, Induce Chromosomal Translocation, and Study DNA Break Repair Mechanisms. mSphere. 2017 Jan 18;2(1). pii: e00340-16. doi: 10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb.	Express gRNA and Cas9 in Leishmania with Neomycin resistance pSP72
	pLdCH	gRNA and Cas9 (Other)	L. donovani ribosome RNA promoter	Matlashewski	Optimized CRISPR-Cas9 Genome Editing for Leishmania and Its Use To Target a Multigene Family, Induce Chromosomal Translocation, and Study DNA Break Repair Mechanisms. mSphere. 2017 Jan 18;2(1). pii: e00340-16. doi: 10.1128/mSphere.00340-16. eCollection 2017 Jan-Feb.	Express gRNA and Cas9 in Leishmania with Hygromycin resistance pSP72
	pJYS3_ΔcrtYf	Cpf1 (Other), crRNA of crtYf (Synthetic)		Yang	CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum. Nat Commun. 2017 May 4;8:15179. doi: 10.1038/ncomms15179.	Constitutive transcription of FnCpf1 and crRNA of crtYf pXMJ19
	pJYS1Ptac	Cpf1 (Other), recT (Other)	tac, unknown	Yang	CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum. Nat Commun. 2017 May 4;8:15179. doi: 10.1038/ncomms15179.	Constitutive trancription of FnCpf1 and recT in C.glutamitum, tac promoter pXMJ19
	pJYS1Peftu	Cpf1 (Other), recT (Other)	etfu, unknown	Yang	CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum. Nat Commun. 2017 May 4;8:15179. doi: 10.1038/ncomms15179.	Constitutive expression of FnCpf1 and recT in C.glutamitum, etfu promoter pXMJ19
	pX2-Cas9	Cas9 (Other)	pBAD	Gill	pX2-Cas9 (unpublished)	Arabinose inducible Cas9 in a broad host range backbone. pBTBX-2
	pSHS212	Cas9 (Other)	T7	Doudna	Conformational control of DNA target cleavage by CRISPR-Cas9. Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 2015 Oct 28.	Cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid pCT10
	pSHS293	Cas9 (Other)	T7	Doudna	Conformational control of DNA target cleavage by CRISPR-Cas9. Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 2015 Oct 28.	D435C/E945C in cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid, lobe closure FRET construct pCT10
	pSHS306 - Bacterial expression plasmid for SpCas9, HNH FRET variant	Cas9 (Other)	T7	Doudna	Conformational control of DNA target cleavage by CRISPR-Cas9. Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 2015 Oct 28.	S867C/S355C in cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid, HNH-1 FRET construct pCT10
	pSHS248	Cas9 (Other)	T7	Doudna	Conformational control of DNA target cleavage by CRISPR-Cas9. Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 2015 Oct 28.	S867C/N1054C in cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid, HNH-2 FRET construct pCT10
	pET-MBP-Geo_st	GeoCas9 (Other)		Doudna	GeoCas9 Plasmids (unpublished)	Expression plasmid for Cas9 from Geobacillus stearothermophilus with an N-Term MBP pET
	pET-MBP-NLS-Geo_st	GeoCas9 (Other)		Doudna	GeoCas9 Plasmids (unpublished)	Expression plasmid for Cas9 from Geobacillus stearothermophilus with an N-Term MBP and SV40 NLS pET
	pFC330	Cas9 (Synthetic), pyrG (Other)	Aspergillus nidulans tef1 promoter, native promoter	Mortensen	A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015.	AMA1 plasmid with Aspergillus optimized Cas9 and pyrG selection marker custom
	pFC331	Cas9 (Synthetic), argB (Other)	Aspergillus nidulans tef1 promoter, native promoter	Mortensen	A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015.	AMA1 plasmid with Aspergillus optimized Cas9 and argB selection marker custom
	pFC333	Cas9 (Synthetic), ble (bleomycin resistance marker) (Other)	Aspergillus nidulans tef1 promoter, Aspergillus nidulans trpC promoter	Mortensen	A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015.	AMA1 plasmid with Aspergillus optimized Cas9 and ble selection marker custom
	pFC332	Cas9 (Synthetic), hph (hygromycin resistance marker) (Other)	Aspergillus nidulans tef1 promoter, Aspergillus nidulans trpC promoter	Mortensen	A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015.	AMA1 plasmid with Aspergillus optimized Cas9 and hph selection marker custom
	pMJ915v2+Nterm Spe1 +Cterm Age1 site			Doudna	Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.	For expression of modified SpyCas9 in e.coli. pMJ915
	1xNLS-pMJ915v2	Cas9		Doudna	Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.	For expression of modified SpyCas9 in e.coli. pMJ915
	2xNLS-pMJ915v2	Cas9, T7		Doudna	Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.	For expression of modified SpyCas9 in e.coli. pMJ915
	4xNLS-pMJ915v2	Cas9, T7		Doudna	Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.	For expression of modified SpyCas9 in e.coli. pMJ915
	pMJ915v2 + sfGFP	Cas9, T7		Doudna	Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.	For expression of modified SpyCas9 in e.coli. pMJ915
	1xNLS-pMJ915v2-sfGFP	Cas9, T7		Doudna	Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.	For expression of modified SpyCas9 in e.coli. pMJ915
	2xNLS-pMJ915v2-sfGFP	Cas9, T7		Doudna	Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.	For expression of modified SpyCas9 in e.coli. pMJ915
	4xNLS-pMJ915v2-sfGFP	Cas9, T7		Doudna	Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.	For expression of modified SpyCas9 in e.coli. pMJ915
	7xNLS-pMJ915v2-sfGFP	Cas9, T7		Doudna	Efficient genome editing in the mouse brain by local delivery of engineered Cas9 ribonucleoprotein complexes. Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.	For expression of modified SpyCas9 in e.coli. pMJ915
	pET-CjCas9	CjCas9 (Other)	T7	Kim	In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni. Nat Commun. 2017 Feb 21;8:14500. doi: 10.1038/ncomms14500.	Expression of CjCas9 with His tag in E.coli pET28b
	pEM-Cas9HF1	Cas9HF1 (Other)	pTet	Lynch	Managing the SOS Response for Enhanced CRISPR-Cas-Based Recombineering in E. coli through Transient Inhibition of Host RecA Activity. ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174.	Modified from pCas9-CR4 (Addgene: 62655) to use the high-fidelity version of Cas9, SpCas9-HF1 (N497A/R661A/Q695A/Q926A) from Kleinstiver et al 2016. pCas9-CR4
	pEM-Cas9HF1-recA56	Cas9HF1 (Other), recA56 (Other)	pTet, proD	Lynch	Managing the SOS Response for Enhanced CRISPR-Cas-Based Recombineering in E. coli through Transient Inhibition of Host RecA Activity. ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174.	Modified from pEM-Cas9HF1 (Addgene ID: 89961) to include constitutive recA56 to block recA-mediated double-strand break repair. pEM-Cas9HF1
	6-His-MBP-TEV-FnCpf1	FnCpf1 (humanized) (Other)	T7	Zhang	Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038.	Bacterial expression plasmid for protein purification pET-28
	6His-MBP-TEV-huAsCpf1	huAsCpf1 (Other)		Zhang	Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5.	Bacterial expression plasmid for protein purification pET-28
	6His-MBP-TEV-huLbCpf1	huLbCpf1 (Other)		Zhang	Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array. Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5.	Bacterial expression plasmid for protein purification pET-28
	pMST665_BB3_L 23_gRNAempty_cas9_BbsI	Cas9		Sauer	An efficient tool for metabolic pathway construction and gene integration for Aspergillus niger. Bioresour Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 10.1016/j.biortech.2017.05.004.	BB3_L 23_gRNAempty_cas9_BbsI; CRISPR plasmid with empty gRNA spacer to incorporate gRNA, Cas9 BB3_AMA_2.8_pUC-ORI_L_AC_hph
	pFREE	Cas9, gRNA array (Synthetic)	ptet, pRhamBAD	Norholm	A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z.	Allows inducible expression of gRNAs and Cas9 for plasmid curing. pMAZ-SK
	pFREE_Amp	Cas9, gRNA array (Synthetic)	ptet, pRhamBAD	Norholm	A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z.	Allows inducible expression of gRNAs and Cas9 for plasmid curing. pMAZ-SK
	pFREE_Cm	Cas9, gRNA array (Synthetic)	ptet, pRhamBAD	Norholm	A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z.	Allows inducible expression of gRNAs and Cas9 for plasmid curing. pMAZ-SK
	pFREE_Zeo	Cas9, gRNA array (Synthetic)	ptet, pRhamBAD	Norholm	A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z.	Allows inducible expression of gRNAs and Cas9 for plasmid curing. pMAZ-SK
	pFREE-RK2	Cas9, gRNA array (Synthetic)	ptet, pRhamBAD	Norholm	A versatile one-step CRISPR-Cas9 based approach to plasmid-curing. Microb Cell Fact. 2017 Aug 2;16(1):135. doi: 10.1186/s12934-017-0748-z.	Allows inducible expression of gRNAs and Cas9 for plasmid curing as well as self-curing via a temperature sensitive RK2 replicon. pMAZ-SK
	pLQ-Pxyl/tet-cas9-Pj23119-sgRNA	Cas9-Pxyltet-sgRNA-pj23119 (Other)		Yang	CRISPR/Cas9-based efficient genome editing in Staphylococcus aureus. Acta Biochim Biophys Sin (Shanghai). 2017 Sep 1;49(9):764-770. doi: 10.1093/abbs/gmx074.	CRISPR-Cas9 based efficient genome editing in S. aureus unknown
	pET28a-Cas9-His	NLS-Cas9-NLS (Other)	T7	Gao	Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes. Nat Commun. 2017 Jan 18;8:14261. doi: 10.1038/ncomms14261.	Purification of Cas9 protein pET28a (+)
	p46Cpf1-OP2	FnCpf1 (Other)	araBAD	Wu	Qiong Wu CRISPR plasmids (unpublished)	Produces lambda Red and Cpf1 for recombination and selection. The plasmid is combined with a donor plasmid (with crRNA and template) for rapid genome editing in E.coli. pKD46
	pJWV102-wtcas9sp	cas9sp (Synthetic)	PczcD	Kjos	Chromosome segregation drives division site selection in Streptococcus pneumoniae. Proc Natl Acad Sci U S A. 2017 Jul 3. pii: 201620608. doi: 10.1073/pnas.1620608114.	Plasmid containing wild-type cas9sp pJWV102
	pThermoCas9_ctrl	Cas9 from the type IIc CRISPR-Cas sytem of Geobacillus thermodenitrificans T12 strain (Other), ThermoCas9 single guide RNA expressing module (Other)	B. smithii xylL promoter, B. coagulans DSM 1 pta promoter	van der Oost	Characterizing a thermostable Cas9 for bacterial genome editing and silencing. Nat Commun. 2017 Nov 21;8(1):1647. doi: 10.1038/s41467-017-01591-4.	Expresses ThermoCas9 and its sgRNA module pNW33n
	SpyCas9(WT)	SpyCas9 (Synthetic)		Huang	Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature. 2017 Jun 15;546(7658):436-439. doi: 10.1038/nature22377. Epub 2017 Apr 27.	Express Streptococcus pyogenes Cas9 pGEX-6P-1 Tag / Fusion Protein GST (N terminal on backbone)
	p6XHis_NLS-SaCas9	CRISPR-associated protein Cas9/Csn1 [Staphylococcus aureus subsp. aureus] (Other)	T7lac	Tarleton	Rapid, Selection-Free, High-Efficiency Genome Editing in Protozoan Parasites Using CRISPR-Cas9 Ribonucleoproteins. MBio. 2017 Nov 7;8(6). pii: e01788-17. doi: 10.1128/mBio.01788-17.	Express SaCas9 in bacteria with a 6xHis tag for purification pET-32 EK/LIC
	pSHS207 - Bacterial expression plasmid for WT SpCas9	SpCas9 (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for WT SpCas9 pCT10
	pJSC033 - Bacterial expression plasmid for SpCas9, REC2 FRET variant	SpCas9 variant C80S/C574S/E60C/D273C (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9, REC2 FRET variant pCT10
	pJSC052 - Bacterial expression plasmid for SpCas9, REC3 FRET variant	SpCas9 variant C80S/C574S/S701C/S960C (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9, REC3 FRET variant pCT10
	pSHS273 - Bacterial expression plasmid for SpCas9∆REC3 variant	SpCas9 variant M1–N497,GGS,V713–D1368 (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9∆REC3 variant pCT10
	pJSC038 - Bacterial expression plasmid for SpCas9∆REC3, HNH FRET variant	SpCas9 variant C80S/C574S/S355C/S867C/M1–N497,GGS,V713–D1368 (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9∆REC3, HNH FRET variant pCT10
	pJSC057 - Bacterial expression plasmid for SpCas9∆REC3, REC2 FRET variant	SpCas9 variant C80S/C574S/E60C/D273C/M1–N497,GGS,V713–D1368 (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9∆REC3, REC2 FRET variant pCT10
	pSHS325 - Bacterial expression plasmid for SpCas9 REC3 domain	SpCas9 variant K506–Q712 (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 REC3 domain pCT10
	pJSC176 - Bacterial expression plasmid for SpCas9 + Q926A variant	SpCas9 variant Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 + Q926A variant pCT10
	pJSC119 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variant	SpCas9 variant N497A/R661A/Q695A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variant pCT10
	pJSC120 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A, HNH FRET variant	SpCas9 variant C80S/C574S/S355C/S867C/N497A/R661A/Q695A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A, HNH FRET variant pCT10
	pJSC111 - Bacterial expression plasmid for SpCas9-HF1 variant	SpCas9 variant N497A/R661A/Q695A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9-HF1 variant pCT10
	pJSC115 - Bacterial expression plasmid for SpCas9-HF1, HNH FRET variant	SpCas9 variant C80S/C574S/S355C/S867C/N497A/R661A/Q695A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9-HF1, HNH FRET variant pCT10
	pJSC129 - Bacterial expression plasmid for SpCas9-HF1, REC2 FRET variant	SpCas9 variant C80S/C574S/E60C/D273C/N497A/R661A/Q695A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9-HF1, REC2 FRET variant pCT10
	pJSC131 - Bacterial expression plasmid for SpCas9-HF1, REC3 FRET variant	SpCas9 variant C80S/C574S/S701C/S960C/N497A/R661A/Q695A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9-HF1, REC3 FRET variant pCT10
	pJSC090 - Bacterial expression plasmid for SpCas9 + K855A variant	SpCas9 variant K855A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 + K855A variant pCT10
	pJSC077 - Bacterial expression plasmid for SpCas9 + K855A, HNH FRET variant	SpCas9 variant C80S/C574S/S355C/S867C/K855A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 + K855A, HNH FRET variant pCT10
	pJSC114 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1) variant	SpCas9 variant K848A/K1003A/R1060A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 eSpCas9(1.1) variant pCT10
	pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant	SpCas9 variant K848A/K1003A/R1060A/N497A/R661A/Q695A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant pCT10
	pJSC173 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9) variant	SpCas9 variant N692A/M694A/Q695A/H698A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9) variant pCT10
	pJSC269 - Bacterial expression plasmid for SpCas9 Cluster 1 + Q926A variant	SpCas9 variant N692A/M694A/Q695A/H698A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 1 + Q926A variant pCT10
	pJSC011 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), HNH FRET variant	SpCas9 variant C80S/C574S/S355C/S867C/N692A/M694A/Q695A/H698A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), HNH FRET variant pCT10
	pJSC281 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC2 FRET variant	SpCas9 variant C80S/C574S/E60C/D273C/N692A/M694A/Q695A/H698A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC2 FRET variant pCT10
	pJSC282 - Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC3 FRET variant	SpCas9 variant C80S/C574S/S701C/S960C/N692A/M694A/Q695A/H698A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9), REC3 FRET variant pCT10
	pJSC197 - Bacterial expression plasmid for SpCas9 Cluster 2 + Q926A variant	SpCas9 variant G528A/V583A/E584A/D585A/N588A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 2 + Q926A variant pCT10
	pJSC228 - Bacterial expression plasmid for SpCas9 Cluster 2 variant	SpCas9 variant G582A/V583A/E584A/D585A/N588A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 2 variant pCT10
	pJSC239 - Bacterial expression plasmid for SpCas9 Cluster 3 + Q926A variant	SpCas9 variant T657A/G658A/W659A/R661A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 3 + Q926A variant pCT10
	pJSC236 - Bacterial expression plasmid for SpCas9 Cluster 4 + Q926A variant	SpCas9 variant F491A/M495A/T496A/N497A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 4 + Q926A variant pCT10
	pJSC272 - Bacterial expression plasmid for SpCas9 Cluster 5 + Q926A variant	SpCas9 variant K918A/V922A/R925A/Q926A (Other)	T7	Doudna	Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature. 2017 Sep 20. doi: 10.1038/nature24268.	Bacterial expression plasmid for SpCas9 Cluster 5 + Q926A variant pCT10
	pEM-Cas9noSsrA	Cas9 (Other)	pTet	Lynch	Managing the SOS Response for Enhanced CRISPR-Cas-Based Recombineering in E. coli through Transient Inhibition of Host RecA Activity. ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174.	Modified from pCas9-CR4 (Addgene: 62655) to remove the ssrA tag from Cas9. pCas9-CR4
	pEM-Cas9HF1-indRecA56	Cas9HF1 (Other), recA56	pTet, pTet	Lynch	Managing the SOS Response for Enhanced CRISPR-Cas-Based Recombineering in E. coli through Transient Inhibition of Host RecA Activity. ACS Synth Biol. 2017 Oct 2. doi: 10.1021/acssynbio.7b00174.	Modified from pEM-Cas9HF1 (Addgene ID: 89961) to co-express inducible recA56 to block recA-mediated double-strand break repair. pEM-Cas9HF1
	AsCpf1-2NLS	AsCpf1-6xHis-MBP-TEV (Synthetic)	T7	Doudna	CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing. Nat Commun. 2017 Dec 8;8(1):2024. doi: 10.1038/s41467-017-01836-2.	bacterial expression of AsCpf1-2NLS pET
	LbCpf1-2NLS	LbCpf1 (Synthetic)	T7	Doudna	CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing. Nat Commun. 2017 Dec 8;8(1):2024. doi: 10.1038/s41467-017-01836-2.	bacterial expression of LbCpf1-2NLS pET
	pSBY1_FnCpf1cg	FnCpf1 (Other)	hsp60	Yang	A CRISPR-Cpf1-Assisted Non-Homologous End Joining Genome Editing System of Mycobacterium smegmatis. Biotechnol J. 2018 Sep;13(9):e1700588. doi: 10.1002/biot.201700588. Epub 2018 Aug 6.	CRISPR system used to genome editing in Mycobacterium smegmatis, expresses Fn CPf1 pMV261
	pJK02	Upstream & Downstream pyrE deletion region (Other), tetR, Cas9 (Other), gRNA		Sorg	Using CRISPR-Cas9-mediated genome editing to generate C. difficile mutants defective in selenoproteins synthesis. Sci Rep. 2017 Nov 7;7(1):14672. doi: 10.1038/s41598-017-15236-5.	Clostridium difficile CRISPR-cas9 mutagenesis plasmid traJ oriT pMTL84151
	pKM126	Upstream & Downstream pyrE deletion region (Other), tetR, Cas9 (Other), gRNA		Sorg	Using CRISPR-Cas9-mediated genome editing to generate C. difficile mutants defective in selenoproteins synthesis. Sci Rep. 2017 Nov 7;7(1):14672. doi: 10.1038/s41598-017-15236-5.	Clostridium difficile CRISPR-cas9 mutagenesis plasmid Tn916 oriT pJS116
	pCas9	cas9 (Other), Lambda red genes (Other)	J23105, araBAD	Pfleger	Genetic tools for reliable gene expression and recombineering in Pseudomonas putida. J Ind Microbiol Biotechnol. 2018 Jan 3. pii: 10.1007/s10295-017-2001-5. doi: 10.1007/s10295-017-2001-5.	Recombineering plasmid with constitutively expressed cas9 and the araBAD promoter expressing αβγ pSEVA224
	p2T-CAG-KKHSaCas9-BlastR	KKH SaCas9	Chicken ß-Actin	Sherwood	Predictable and precise template-free CRISPR editing of pathogenic variants. Nature. 2018 Nov;563(7733):646-651. doi: 10.1038/s41586-018-0686-x. Epub 2018 Nov 7.	Confers constitutive expression of KKH SaCas9. p2T-CAG-MCS-BlastR NEWENTRY
	pCAS9counter	Cas9, sgRNA	prab17, prab17	Salipante	Efficient and Scalable Precision Genome Editing in Staphylococcus aureus through Conditional Recombineering and CRISPR/Cas9-Mediated Counterselection. MBio. 2018 Feb 20;9(1). pii: mBio.00067-18. doi: 10.1128/mBio.00067-18.	Expresses CAS9 and sgRNA for targeted counterselection in S. aureus pCN-50
	pET28b-NmCas9-Flag	CRISPR-associated endonuclease Cas9 (Other)	T7	Zhang	Programmable RNA Cleavage and Recognition by a Natural CRISPR-Cas9 System from Neisseria meningitidis. Mol Cell. 2018 Mar 1;69(5):906-914.e4. doi: 10.1016/j.molcel.2018.01.025. Epub 2018 Feb 15.	Wild type Cas9 from Nm8013 cloned into pET28b with a C-terminal Flag tag for protein expression pET28b
	pxCas9CR4	TetR and xCas9 (Synthetic)		Reisch	pCas9CR4 with point mutations from Cas9X for NG PAM sites (unpublished)	PCas9CR4 with the xCas9 mutations allowing NG PAM sites p15A origin
	pRPaCas9	Cas9 (Other)		Horn	Inducible high-efficiency CRISPR-Cas9-targeted gene editing and precision base editing in African trypanosomes. Sci Rep. 2018 May 21;8(1):7960. doi: 10.1038/s41598-018-26303-w.	Expresses Cas9 in T. brucei (2T1) cells pUC
	pCas9-J23109	Cas9 (Other)		Zhang	Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572.	Plasmid constitutively expressing Cas9 protein N.A.
	peSpCas9-J23109	eSpCas9 (Other)		Zhang	Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572.	Plasmid constitutively expressing eSpCas9 protein N.A.
	pCasPA			Ji	CRISPR/Cas9-based Genome Editing in Pseudomonas aeruginosa and Cytidine Deaminase-Mediated Base Editing in Pseudomonas Species. iScience. 2018 Aug 31;6:222-231. doi: 10.1016/j.isci.2018.07.024. Epub 2018 Aug 1.	Bacterial expression of Cas9 nuclease and λ-Red system in Pseudomonas aeruginosa unknown
	pNS20-SpCas9-SNAP	Cas9 (Other)		Schwank	Covalent linkage of the DNA repair template to the CRISPR-Cas9 nuclease enhances homology-directed repair. Elife. 2018 May 29;7. pii: 33761. doi: 10.7554/eLife.33761.	Bacterial vector for expression of Snap-tagged Streptococcus pyogenes Cas9 pET His6 MBP N10 TEV LIC cloning vector (2C-T)
	pET-21a_3xNLS_SpCas9_protein_expression	3xNLS_SpCas9 (Synthetic)	T7	Wolfe	Highly efficient therapeutic gene editing of human hematopoietic stem cells. Nat Med. 2019 May;25(5):776-783. doi: 10.1038/s41591-019-0401-y. Epub 2019 Mar 25.	Protein expression plasmid for 3xNLS SpCas9 in pET-21a backbone pET-21 a
	pET-21a_2xNLS_LbCpf1_protein_expression	2xNLS_LbCpf1 (Synthetic)	T7	Wolfe	Editing aberrant splice sites efficiently restores beta-globin expression in beta-thalassemia. Blood. 2019 Jan 31. pii: blood-2019-01-895094. doi: 10.1182/blood-2019-01-895094.	Protein expression plasmid for 2xNLS LbCpf1 in pET-21a backbone pET-21 a
	pJL1-SpCas9	S. pyogenes Cas9 (Other)	T7	Jewett	BioBits Health: Classroom Activities Exploring Engineering, Biology, and Human Health with Fluorescent Readouts. ACS Synth Biol. 2019 May 7. doi: 10.1021/acssynbio.8b00381.	In vitro expression of S. pyogenes Cas9 from the T7 promoter pJL1
	pCasKP-apr	Cas9 (Other)		Ji	Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18.	Bacterial expression of Cas9 nuclease and lambda-Red system in Klebsiella Pneumoniae Unknown
	pCasKP-hph	Cas9 (Other)		Ji	Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18.	Bacterial expression of Cas9 nuclease and lambda-Red system in Klebsiella Pneumoniae Unknown
	pHSB04X	Cas9, promotor PslpA-guide-RNA, homologous arms of Lb_1019 (Other)		Yang	Development of a RecE/T-Assisted CRISPR-Cas9 Toolbox for Lactobacillus. Biotechnol J. 2019 Jul;14(7):e1800690. doi: 10.1002/biot.201800690. Epub 2019 May 20.	Genome editing for Lactobacillus brevis ATCC367 pCas
	pHSP02	Cas9, promotor P11-guide-RNA, homologous arms of Lp_0537 (Other)		Yang	Development of a RecE/T-Assisted CRISPR-Cas9 Toolbox for Lactobacillus. Biotechnol J. 2019 Jul;14(7):e1800690. doi: 10.1002/biot.201800690. Epub 2019 May 20.	Genome editing for Lactobacillus plantarum WCSF1 pCas
	p15A_SpyCas9_CmR	Cas9 from S. pyogenes		Noireaux	An educational module to explore CRISPR technologies with a cell-free transcription-translation system (unpublished)	Expresses S. pyogenes Cas9 p15A, Chloramphenicol resistance NEWENTRY
	pCas9-J23113	Cas9 (Other)		Zhang	Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572.	Expression of Cas9 with a weak promoter p15A origin
	Nme2Cas9_pMCSG7	Nme2Cas9 (Other)	T7	Sontheimer	A Compact, High-Accuracy Cas9 with a Dinucleotide PAM for In Vivo Genome Editing. Mol Cell. 2018 Dec 18. pii: S1097-2765(18)31033-5. doi: 10.1016/j.molcel.2018.12.003.	Nme2Cas9 bacterial expression plasmid pMCSG7
	NmeCas9_3XNLS_pMCSG7	NmeCas9 (Other)		Sontheimer	NmeCas9 is an intrinsically high-fidelity genome editing platform (unpublished)	Bacterial expression of wtNmeCas9 with 3X NLS pMCSG7
	pCas9CR4-VQR	SpCas9-VQR (Synthetic)	pTet	Reisch	pCas9CR4 variants for increased targeting space (unpublished)	pCas9CR4 with VQR mutations for changing PAM site specificity to NGAN or NGNG from Kleinstiver et al. 2015 pdCas9-bacteria
	pCas9CR4-NG	SpCas9-NG (Synthetic)	pTet	Reisch	pCas9CR4 variants for increased targeting space (unpublished)	pCas9CR4 with mutations to change PAM site specificity to NG from Nishimasu et al. pdCas9-bacteria
	pCas9CR5	SpCas9 (Synthetic)	pTet	Reisch	pCas9CR4 variants for increased targeting space (unpublished)	pCas9CR4 with stop codon to prevent translation of ssrA degradation tag pdCas9-bacteria
	pCas9CR5-NG	SpCas9-NG (Synthetic)	pTet	Reisch	pCas9CR4 variants for increased targeting space (unpublished)	pCas9CR5 with mutations to change PAM site specificity to NG from Nishimasu et al. pdCas9-bacteria
	pJZ002			Moore	Refactoring the Cryptic Streptophenazine Biosynthetic Gene Cluster Unites Phenazine, Polyketide, and Nonribosomal Peptide Biochemistry. Cell Chem Biol. 2019 Feb 15. pii: S2451-9456(19)30038-8. doi: 10.1016/j.chembiol.2019.02.004.	CRISPR-cas9 targeting in E. coli with ampicillin resistance N/A
	pSU-araC-Cas9	Cas9 (Other)	araBAD	Yang	Multigene editing in the Escherichia coli genome using the CRISPR-Cas9 system. Appl Environ Microbiol. 2015 Jan 30. pii: AEM.04023-14.	Multigene editing in the Escherichia coli genome using the CRISPR-Cas9 system p15A
	pET-21a_2xNLS_FnCpf1_MBP_protein_expression	2xNLS_FnCpf1 (Synthetic)	T7	Wolfe	Enhanced Cas12a editing in mammalian cells and zebrafish. Nucleic Acids Res. 2019 Mar 20. pii: 5403491. doi: 10.1093/nar/gkz184.	Protein expression plasmid for 2xNLS FnCpf1 in pET-21a backbone pET-21 a
	pUC-fFuCas9-HTBNLS-hph	Cas9 and hygromycin resist gene (Other)	gpdA;trpC	Ji	CRISPR/Cas9-Based Genome Editing in the Filamentous Fungus Fusarium fujikuroi and Its Application in Strain Engineering for Gibberellic Acid Production. ACS Synth Biol. 2019 Feb 15;8(2):445-454. doi: 10.1021/acssynbio.8b00478. Epub 2019 Jan 23.	Multigene editing in the Fusarium fujikuroi genome using the CRISPR-Cas9 system pUC57
	pEJS1026-pMCSG7-HpaCas9	HpaCas9 (Other)		Sontheimer	Potent Cas9 Inhibition in Bacterial and Human Cells by AcrIIC4 and AcrIIC5 Anti-CRISPR Proteins. MBio. 2018 Dec 4;9(6). pii: mBio.02321-18. doi: 10.1128/mBio.02321-18.	Expresses a 6X-His tagged type II-C Cas9 from H. parainfluenzae in bacterial cells pMCSG7
	pEJS1027-pMCSG7-SmuCas9	SmuCas9 (Other)		Sontheimer	Potent Cas9 Inhibition in Bacterial and Human Cells by AcrIIC4 and AcrIIC5 Anti-CRISPR Proteins. MBio. 2018 Dec 4;9(6). pii: mBio.02321-18. doi: 10.1128/mBio.02321-18.	Expresses a 6X-His tagged type II-C Cas9 from S. muelleri in bacterial cells pMCSG7
	pCasAb-apr			Ji	A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of OxyR. Cell Chem Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi: 10.1016/j.chembiol.2019.09.003.	Bacterial expression of Cas9 nuclease and RecAb recombination system in Acinetobacter baumannii pMMB67EH
	pCpf1			Zhang	Expanding the potential of CRISPR-Cpf1 based genome editing technology in the cyanobacterium Anabaena PCC 7120. ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437.	Bacterial genome editing plasmid with kanamycin resistance marker Unknown
	pCpf1-sp			Zhang	Expanding the potential of CRISPR-Cpf1 based genome editing technology in the cyanobacterium Anabaena PCC 7120. ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437.	Bacterial genome editing plasmid with spectinomycin resistance marker Unknown
	pCpf1b			Zhang	Expanding the potential of CRISPR-Cpf1 based genome editing technology in the cyanobacterium Anabaena PCC 7120. ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437.	Bacterial genome editing plasmid with kanamycin resistance marker and using sacB as a counter-selection marker Unknown
	pCpf1b-sp			Zhang	Expanding the potential of CRISPR-Cpf1 based genome editing technology in the cyanobacterium Anabaena PCC 7120. ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437.	Bacterial genome editing plasmid with spectinomycin resistance marker and using sacB as a counter-selection marker Unknown
	pLdSaCN	gRNA and SaCas9 (Other)		Matlashewski	Single-Strand Annealing Plays a Major Role in Double-Strand DNA Break Repair following CRISPR-Cas9 Cleavage in Leishmania. mSphere. 2019 Aug 21;4(4). pii: 4/4/e00408-19. doi: 10.1128/mSphere.00408-19.	Expresses Staphylococcus aureus Cas9 (SaCas9) and its gRNA in Leishmania pSP72
	pLPhygSaCas9	Staphylococcus aureus Cas9 (Other)		Matlashewski	Single-Strand Annealing Plays a Major Role in Double-Strand DNA Break Repair following CRISPR-Cas9 Cleavage in Leishmania. mSphere. 2019 Aug 21;4(4). pii: 4/4/e00408-19. doi: 10.1128/mSphere.00408-19.	Expresses Staphylococcus aureus Cas9 (SaCas9) in Leishmania pSP72 NEWENTRY
	PCV-Cas9	PCV2 (Other), Cas9 (Other)		Gordon	Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template. Commun Biol. 2018 May 31;1:54. doi: 10.1038/s42003-018-0054-2. eCollection 2018.	Expresses Cas9 with an N-terminal HUH-tag called PCV2 pTD68 ORF1 PCV2_gp1
	Cas9-PCV	PCV2 (Other), Cas9 (Other)		Gordon	Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template. Commun Biol. 2018 May 31;1:54. doi: 10.1038/s42003-018-0054-2. eCollection 2018.	Expresses Cas9 with a C-terminal HUH-tag called PCV2 in E. coli pTD68 ORF1 PCV2_gp1
	spCas9-NLS-MAV	cas9 (Other)	T7	Akavia	Double-Stranded Biotinylated Donor Enhances Homology-Directed Repair in Combination with Cas9 Monoavidin in Mammalian Cells. CRISPR J. 2018 Dec;1:414-430. doi: 10.1089/crispr.2018.0045.	SpCas9-NLS 17 amino acid linker to monoavidin (SpCas9-NLS-MAV) pEC-K-MBP
	pET-21a_FnCpf1_2xNLS_protein_expression	FnCpf1_2xNLS (Synthetic)		Wolfe	Enhanced Cas12a editing in mammalian cells and zebrafish. Nucleic Acids Res. 2019 Mar 20. pii: 5403491. doi: 10.1093/nar/gkz184.	Protein expression plasmid for 2xNLS FnCpf1 in pET-21a backbone pET-21 a
	pFD115	Ptet (Other), cas9 (Other), term PpflB sgRNA (BsaI) (Synthetic)	Ptet	Bikard	Gene silencing with CRISPRi in bacteria and optimization of dCas9 expression levels. Methods. 2019 Aug 1. pii: S1046-2023(18)30497-3. doi: 10.1016/j.ymeth.2019.07.024.	pFD115 carries cas9 controlled by an aTc-inducible Ptet promoter, a sgRNA controlled constitutively from PpflB S. aureus promoter to clone a guide between two BsaI sites and oriT on pLZ12 vector pLZ12
	pCold CL7-Cas9	Cas9 (Other)		Liu	Co-expression of Cas9 and single-guided RNAs in Escherichia coli streamlines production of Cas9 ribonucleoproteins. Commun Biol. 2019 May 3;2:161. doi: 10.1038/s42003-019-0402-x. eCollection 2019.	obtaining full Cas9 RNP pCold I
	pLdCN2	gRNA and Cas9 (Other)		Matlashewski	Application of CRISPR/Cas9-Mediated Genome Editing in Leishmania. Methods Mol Biol. 2020;2116:199-224. doi: 10.1007/978-1-0716-0294-2_14.	Expresses Streptococcus pyogenes Cas9 (SpCas9) and two gRNAs in Leishmania pLdCN
	pLQ-KO-tgt50	Cas9-Pxyltet-sgRNA-Pspac-donor-tgt50 (Other)		Yang	CRISPR/Cas9-based efficient genome editing in Staphylococcus aureus. Acta Biochim Biophys Sin (Shanghai). 2017 Sep 1;49(9):764-770. doi: 10.1093/abbs/gmx074.	CRISPR-Cas9 based efficient genome editing in S. aureus ColE1 origin, AmpR, Rep(+), cat resistance
	pET-FLAG-eSpCas9	eSpCas9 (Other)	T7	Welker	Blackjack mutations improve the on-target activities of all increased fidelity variants of SpCas9 without compromising their fidelities Nature Communications, 1223, 2020	Expression of increased fidelity eSpCas9 in bacterial cells pMJ806-like
	pET-FLAG-SpCas9-HF1	SpCas9-HF1 (Other)	T7	Welker	Blackjack mutations improve the on-target activities of all increased fidelity variants of SpCas9 without compromising their fidelities Nature Communications, 1223, 2020	Expression of increased fidelity SpCas9-HF1 in bacterial cells pMJ806-like
	pET-FLAG-B-eSpCas9	B-eSpCas9 (Other)	T7	Welker	Blackjack mutations improve the on-target activities of all increased fidelity variants of SpCas9 without compromising their fidelities Nature Communications, 1223, 2020	Expression of increased fidelity Blackjack-eSpCas9 in bacterial cells. Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with higher fidelity than that of eSpCas9 pMJ806-like
	pET-FLAG-eSpCas9-plus	eSpCas9-plus (Other)	T7	Welker	Blackjack mutations improve the on-target activities of all increased fidelity variants of SpCas9 without compromising their fidelities Nature Communications, 1223, 2020	Expression of increased fidelity eSpCas9-plus in bacterial cells. Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with close to the same fidelity as eSpCas9. pMJ806-like
	pET-FLAG-SpCas9-HF1-plus	SpCas9-HF1-plus (Other)	T7	Welker	Blackjack mutations improve the on-target activities of all increased fidelity variants of SpCas9 without compromising their fidelities Nature Communications, 1223, 2020	Expression of increased fidelity SpCas9-HF1-plus in bacterial cells. Cleaves both 20nt and 5'-extended 21nt spacer containing sgRNAs with close to the same fidelity as SpCas9-HF1. pMJ806-like
	pIgnaviCas9	IgnaviCas9 (Other)	T7 promoter	Quake	Nucleic acid cleavage with a hyperthermophilic Cas9 from an uncultured Ignavibacterium. Proc Natl Acad Sci U S A. 2019 Oct 28. pii: 1904273116. doi: 10.1073/pnas.1904273116.	Expresses IgnaviCas9 in E. coli pMJ806
	pCAH01Sp::Cas9	Sp Cas9 (Other)	Ptet	Henard	Development of a CRISPR/Cas9 System for Methylococcus capsulatus In Vivo Gene Editing. Appl Environ Microbiol. 2019 May 16;85(11). pii: AEM.00340-19. doi: 10.1128/AEM.00340-19. Print 2019 Jun 1.	Ptet-controlled expression of S. pyogenes Cas9 pCAH01SpR
	pCasTet-λ	TetR and pTetR/TetO (Other)	tet promoter	Johnston	Systematic evasion of the restriction-modification barrier in bacteria. Proc Natl Acad Sci U S A. 2019 May 16. pii: 1820256116. doi: 10.1073/pnas.1820256116.	Constitutive expression of cas9 and anhydrotetracycline/tetracycline inducible expression of lamda RED. Useful variant of Plasmid #62225 when arabinose induction is not possible. Plasmid #62225
	pSDMA65	Hygromycin (HYG) (Synthetic)	ACT1	Fraser	Targeted Genome Editing via CRISPR in the Pathogen Cryptococcus neoformans. (unpublished)	Codon optimised Streptococcus pyogenes CAS9 ORF regulated by the Cryptococcus neoformans TEF1 promoter and terminator. pBlueScript II SK(-)
	pCRISPomyces-Sth1Cas9	Sth1Cas9 (Other)	rpsL(XC)-BbsI	Wong	Characterization of Cas proteins for CRISPR-Cas editing in streptomycetes. Biotechnol Bioeng. 2019 May 15. doi: 10.1002/bit.27021.	Contains codon-optimized Sth1Cas9 and gRNA for streptomyces pKC1139
	pCRISPomyces-SaCas9	SaCas9 (Other)	rpsL(XC)-BbsI	Wong	Characterization of Cas proteins for CRISPR-Cas editing in streptomycetes. Biotechnol Bioeng. 2019 May 15. doi: 10.1002/bit.27021.	Contains codon-optimized SaCas9 and gRNA for streptomyces pKC1139
	pCRISPomyces-FnCpf1	FnCpf1 (Other)	rpsL(XC)-BbsI	Wong	Characterization of Cas proteins for CRISPR-Cas editing in streptomycetes. Biotechnol Bioeng. 2019 May 15. doi: 10.1002/bit.27021.	Contains codon-optimized FnCpf1 and gRNA for streptomyces pKC1139
	pET-His6-FnCas9GFP	HA-NLS-FnCas9 (Synthetic)	T7	Chakraborty	Francisella novicida Cas9 interrogates genomic DNA with very high specificity and can be used for mammalian genome editing. Proc Natl Acad Sci U S A. 2019 Oct 15;116(42):20959-20968. doi: 10.1073/pnas.1818461116. Epub 2019 Sep 30.	expression of FnCas9-GFP in bacterial cells pET-His6-GFP-TEV-LIC
	pKM197	Upstream & Downstream pyrE deletion region (Other)		Sorg	Modified / Updated Clostridium difficile CRISPR-Cas9 genome editing plasmids (unpublished)	pyrE-targeted CRISPR-Cas9 control plasmid. Cas9 expression is controlled by the xylR promoter and results in improved conjugation efficiency. Induction with xylose results in a pyrE mutant. pJS116
	pSCBE3-HF	HF-BE3 (Other)		Sun	Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137	Expresses HF-BE3 CRISPR base editor and gRNA scaffold in Streptomyces pYH7
	pSEVA421-Cas9tr	tracrRNA-Cas9 (Other)		de Lorenzo	Recombination-Independent Genome Editing through CRISPR/Cas9-Enhanced TargeTron Delivery. ACS Synth Biol. 2019 Sep 20;8(9):2186-2193. doi: 10.1021/acssynbio.9b00293. Epub 2019 Sep 3.	Plasmid constitutively expressing the Cas9 nuclease and the non-coding tracrRNA from S. pyogenes pSEVA421
	pCRISPRx-Sth1Cas9-L5	Sth1Cas9 (Other), TetR, L5 attP, L5 Int, KanR		Bitter	Efficient genome editing in pathogenic mycobacteria using Streptococcus thermophilus CRISPR1-Cas9 Tuberculosis (2020) 101983	Sth1Cas9, TetR, KanR, L5Int, attP to create frameshifts and gene deletion in Mycobacteria PLJR962
	pCB578	Cas9 and tracrRNA (Other), repeat-spacer-repeat array		Beisel	Genome Editing with CRISPR-Cas9 in Lactobacillus plantarum Revealed That Editing Outcomes Can Vary Across Strains and Between Methods. Biotechnol J. 2019 Mar;14(3):e1700583. doi: 10.1002/biot.201700583. Epub 2018 Sep 20.	E. coli-Lactobacilli shuttle vector containing SpCas9, tracrRNA, and a repeat-spacer-repeat array colE1, pAM beta
	pCgCas9_recT	Cas9 (Other)		Yang	De Novo Engineering of Corynebacterium glutamicum for l-Proline Production. ACS Synth Biol. 2020 Jul 17;9(7):1897-1906. doi: 10.1021/acssynbio.0c00249. Epub 2020 Jul 6.	Double-plasmid-based CRISPR-Cas9 system in Corynebacterium glutamicum; pBL1ts oriVC. glutamicum; pSC101 oriVE. coli PlacM-SpCas9 Streptomyces coelicolor codon-optimized, Peftu-RecT; Kanr pJYS1Peftu
	pJH1			Poulter	The application of the CRISPR-Cas9 system in Pseudomonas syringae pv. actinidiae. J Med Microbiol. 2020 Mar;69(3):478-486. doi: 10.1099/jmm.0.001124. Epub 2020 Jan 14.	encodes CRISPR-Cas9 system derived from plasmid pCas9 pCas9

Base Edit

Catalytically dead dCas9 fused to a cytidine deaminase protein becomes a specific cytosine base editor that can alter DNA bases without inducing a DNA break. Cytosine base editors convert C->T (or G->A on the opposite strand) within a small editing window specified by the gRNA. Adenine base editors convert adenine to inosine, which is replaced by guanosine to create A->G (or T->C on the opposite strand) mutations.

	Plasmid	Promoter	PI	Publication	Hidden Extra Search Info
	pET42b-BE3	T7	Liu	Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery. Nat Commun. 2017 Jun 6;8:15790. doi: 10.1038/ncomms15790.	Expresses BE3-NLS with an N-terminal His Tag (His6) for bacterial expression pET_42b
	pET42b-HF-BE3		Liu	Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery. Nat Commun. 2017 Jun 6;8:15790. doi: 10.1038/ncomms15790.	Expresses HF-BE3-NLS with an N-terminal His Tag (His6) for bacterial expression pET_42b
	pET28b-BE3∆UGI	T7	Kim	Genome-wide target specificities of CRISPR RNA-guided programmable deaminases. Nat Biotechnol. 2017 May;35(5):475-480. doi: 10.1038/nbt.3852. Epub 2017 Apr 10.	The plasmid encoding the His6-rAPOBEC1-XTEN-nCas9 protein (BE3∆UGI) was generated by site-directed mutagenesis using pET28b-BE1 (Addgene plasmid #73018). pET28b
	pScI_dCas9-CDA	lambda OR	Kondo	Deaminase-mediated multiplex genome editing in Escherichia coli. Nat Microbiol. 2018 Feb 5. pii: 10.1038/s41564-017-0102-6. doi: 10.1038/s41564-017-0102-6.	Bacterial Target-AID vector pSC101
	pScI_dCas9-CDA_J23119-sgRNA	lambda OR, J23119	Kondo	Deaminase-mediated multiplex genome editing in Escherichia coli. Nat Microbiol. 2018 Feb 5. pii: 10.1038/s41564-017-0102-6. doi: 10.1038/s41564-017-0102-6.	Bacterial Target-AID vector with sgRNA pSC101
	pScI_dCas9-CDA-UL	lambda OR	Kondo	Deaminase-mediated multiplex genome editing in Escherichia coli. Nat Microbiol. 2018 Feb 5. pii: 10.1038/s41564-017-0102-6. doi: 10.1038/s41564-017-0102-6.	Bacterial Target-AID UGI-LVA vector pSC101
	pnCasPA-BEC		Ji	CRISPR/Cas9-based Genome Editing in Pseudomonas aeruginosa and Cytidine Deaminase-Mediated Base Editing in Pseudomonas Species. iScience. 2018 Aug 31;6:222-231. doi: 10.1016/j.isci.2018.07.024. Epub 2018 Aug 1.	A cytidine deaminase-mediated base-editing plasmid in Pseudomonas species unknown
	pBECKP-km		Ji	Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18.	A cytidine deaminase-mediated base-editing plasmid in Klebsiella Pneumoniae Unknown
	pBECKP-spe		Ji	Precise and efficient genome editing in Klebsiella pneumoniae using CRISPR-Cas9 and CRISPR-assisted cytidine deaminase. Appl Environ Microbiol. 2018 Sep 14. pii: AEM.01834-18. doi: 10.1128/AEM.01834-18.	ncas9-BEC, fusion protein of rAPOBEC1 and spCas9 nickase Unknown
	pET42b-A3A-PBE-ΔUGI	T7 promoter	Gao	Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A. Nat Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261.	To express the recombinant APOBEC3A in vitro pET42b Tags / Fusion Proteins 6*HIS (N terminal on insert) NLS (C terminal on insert)
	pET42b-ABE7.10	T7	Huang	Genome-wide profiling of adenine base editor specificity by EndoV-seq. Nat Commun. 2019 Jan 8;10(1):67. doi: 10.1038/s41467-018-07988-z.	Expresses ABE7.10-NLS with an N-terminal His Tag (His6) for bacterial expression pET42b
	pBECAb-apr		Ji	A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of OxyR. Cell Chem Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi: 10.1016/j.chembiol.2019.09.003.	A cytidine deaminase-mediated base-editing plasmid in Acinetobacter baumannii pET28a and pWH1266
	pCRISPR-cBEST	ermE*/tipA	Weber	Highly efficient DSB-free base editing for streptomycetes with CRISPR-BEST. Proc Natl Acad Sci U S A. 2019 Oct 8;116(41):20366-20375. doi: 10.1073/pnas.1913493116. Epub 2019 Sep 23.	C to T base editor for actinomycetes pGM1190
	pCRISPR-aBEST	tipA, tipA	Weber	Highly efficient DSB-free base editing for streptomycetes with CRISPR-BEST. Proc Natl Acad Sci U S A. 2019 Oct 8;116(41):20366-20375. doi: 10.1073/pnas.1913493116. Epub 2019 Sep 23.	Expression of a Streptomyces codon optimized Cas9n-tadA fusion protein in pGM1190. A to G base editor for actinomycetes pGM1190
	pSCBE2		Sun	Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137	Expresses BE2 CRISPR base editor and gRNA scaffold in Streptomyces pYH7
	pSCBE3		Sun	Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137	Expresses BE3 CRISPR base editor and gRNA scaffold in Streptomyces pYH7
	pSCBE3-HF		Sun	Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137	Expresses HF-BE3 CRISPR base editor and gRNA scaffold in Streptomyces pYH7
	pSABEd		Sun	Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137	Expresses ABEd CRISPR base editor and gRNA scaffold in Streptomyces pYH7
	pSABEn		Sun	Base editing in Streptomyces with Cas9-deaminase fusions BioRxiv 630137	Expresses ABEn CRISPR base editor and gRNA scaffold in Streptomyces pYH7

Nick

CRISPR/Cas nickase mutants introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type Cas enzymes. To use a nickase mutant, you will need two gRNAs that target opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB) that is repaired using error-prone non-homologous end joining (NHEJ). Double nicking strategies reduce unwanted off-target effects. Nickase mutants can also be used with a repair template to introduce specific edits via homology-directed repair (HDR).

	Plasmid	Gene/Insert	Promoter	PI	Publication	Hidden Extra Search Info
	pMJ825	Cas9	T7	Doudna	A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012 Jun 28.	pEC-K-MBP Tag / Fusion Protein His6 (N terminal on backbone)
	pMJ826	Cas9	T7	Doudna	A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012 Jun 28.	pEC-K-MBP Tag / Fusion Protein His6 (N terminal on backbone)
	pMCSG7-D16A-NmeCas9	pMCSG7-D16A-NmeCas9 (Other)		Sontheimer	DNase H Activity of Neisseria meningitidis Cas9. Mol Cell. 2015 Oct 15;60(2):242-55. doi: 10.1016/j.molcel.2015.09.020.	bacterial pMCSG7 expression vector expressing Nme cas9 nickase with a D16A mutation (RuvC domain) with T7 promoter, N-terminal His tag and TEV site pMCSG7
	pMCSG7-H588A-NmeCas9	pMCSG7-H588A-NmeCas9 (Other)	T7	Sontheimer	DNase H Activity of Neisseria meningitidis Cas9. Mol Cell. 2015 Oct 15;60(2):242-55. doi: 10.1016/j.molcel.2015.09.020.	bacterial pMCSG7 expression vector expressing Nme cas9 nickase with H588A mutation (HNH domain) with T7 promoter, N-terminal His tag and TEV site pMCSG7
	pNICKclos2.0	Cas9 nickase (Other), sgRNA to xylR (Synthetic)	Pthl, Pj23119	Yang	CRISPR-based genome editing and expression control systems in Clostridium acetobutylicum and Clostridium beijerinckii. Biotechnol J. 2016 May 23. doi: 10.1002/biot.201600053.	Genome editing for gene xylR (cbei-2385) in clostridium beijerinckii NCIMB 8052 pXY1
	pNICKclos1.0	Cas9 nickase (Other), sGRNA to pyrE	ptb, j23119	Yang	CRISPR-based genome editing and expression control systems in Clostridium acetobutylicum and Clostridium beijerinckii. Biotechnol J. 2016 May 23. doi: 10.1002/biot.201600053.	Genome editing for gene pyrE (CAC-002) in Clostridium acetobutylicum ATCC 824 pIMP1-ptb
	pCas9(D10A)	Cas9 D10A (Synthetic)	pTeto	Wang	Targeted Large-Scale Deletion of Bacterial Genomes Using CRISPR-Nickases. ACS Synth Biol. 2015 Nov 20;4(11):1217-25. doi: 10.1021/acssynbio.5b00132. Epub 2015 Oct 25.	Nicking Cas9 derived from pWT Cas9 (Addgene #44250). Ampicillin resistance marker, the tetracycline repressor (TetR), a ColE1 origin of replication and Streptococcus pyogenes Cas9 D10A pWTCas9
	pLCNICK	Cas9 nickase (Other), sgRNA, homology arms of LC2W_2179		Yang	CRISPR-Cas9D10A Nickase-Assisted Genome Editing in Lactobacillus casei. Appl Environ Microbiol. 2017 Sep 1. pii: AEM.01259-17. doi: 10.1128/AEM.01259-17.	Genome editing for Lactobacillus casei Lc2W pCas

Activate

Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest. If the plasmid that you choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator to your specific locus.

	Plasmid	Gene/Insert	Promoter	PI	Publication	Hidden Extra Search Info
	pWJ66	tracrRNA (Other), dcas9-w (Other), CRISPR array		Marraffini	Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic Acids Res. 2013 Jun 12.	Same as pdCas9, but with the dCas9-w fusion (w is fused at the C-terminal end of dCas9) pACYC184
	pWJ68	tracrRNA (Other), w-dcas9 (Other), CRISPR array		Marraffini	Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic Acids Res. 2013 Jun 12.	Same as pdCas9, but with the w-dCas9 fusion (w is fused at the N-terminal end of dCas9) pACYC184
	pET-dCas9-VP64-6xHis	dCas9-VP64 (Other)	T7	Liu	Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 2014 Oct 30.	Expression of dCas9-VP64-6xHis in bacterial cells pET29
	pET-deSpCas9-VP64-6xHis	dead/inactive eSpCas9-NLS-3xFLAG-VP64 (Other)	T7	Welker	Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage. Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8.	Expression of dead/inactive increased fidelity eSpCas9 (1.1)-VP64-6xHis in bacterial cells pET29
	pET-dSpCas9-HF1-VP64-6xHis	dead/inactive SpCas9-HF1-NLS-3xFLAG-VP64 (Other)	T7	Welker	Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage. Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8.	Expression of dead/inactive increased fidelity SpCas9-HF1-VP64-6xHis in bacterial cells pET29
	pET-dHeFSpCas9-VP64-6xHis	dead/inactive HeFSpCas9-NLS-3xFLAG-VP64 (Other)	T7	Welker	Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage. Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8.	Expression of dead/inactive increased fidelity HeFSpCas9-VP64-6xHis in bacterial cells pET29
	pCD185	dCas9 and MCP-SoxS_R93A (Other)		Zalatan	Synthetic CRISPR-Cas gene activators for transcriptional reprogramming in bacteria. Nat Commun. 2018 Jun 27;9(1):2489. doi: 10.1038/s41467-018-04901-6.	dCas9, MCP-SoxS_R93A p15A vector
	pCD227	dCas9 and MCP-SoxS_R93A (Other)		Zalatan	Synthetic CRISPR-Cas gene activators for transcriptional reprogramming in bacteria. Nat Commun. 2018 Jun 27;9(1):2489. doi: 10.1038/s41467-018-04901-6.	AraC-pBAD-dCas9, MCP-SoxS_R93A p15A vector
	pJF093	dCas9 and MCP-SoxS_R93A (Other)		Zalatan	Synthetic CRISPR-Cas gene activators for transcriptional reprogramming in bacteria. Nat Commun. 2018 Jun 27;9(1):2489. doi: 10.1038/s41467-018-04901-6.	Sp.pCas9-dCas9, pTet-MCP-SoxS_R93A p15A vector
	pJF104B	dCas9 and MCP-SoxS_R93A (Other)		Zalatan	Synthetic CRISPR-Cas gene activators for transcriptional reprogramming in bacteria. Nat Commun. 2018 Jun 27;9(1):2489. doi: 10.1038/s41467-018-04901-6.	pTet-dCas9, pTet-MCP-SoxS_R93A p15A vector
	pdCas9-omega	dCas9-omega (Other)	Tet promoter	Chatterjee	Multiplexed deactivated CRISPR-Cas9 gene expression perturbations deter bacterial adaptation by inducing negative epistasis. Commun Biol. 2018 Sep 3;1:129. doi: 10.1038/s42003-018-0135-2. eCollection 2018.	Addgene plasmid 44249 where dCas9 has been replaced with dCas9 fused to the omega subunit of RNAP p15a
	pFD116	Ptet (Other), dcas9 (Other), PpflB sgRNA BsaI (Synthetic)		Bikard	Gene silencing with CRISPRi in bacteria and optimization of dCas9 expression levels. Methods. 2019 Aug 1. pii: S1046-2023(18)30497-3. doi: 10.1016/j.ymeth.2019.07.024.	pFD116 carries dcas9 controlled by an aTc-inducible Ptet promoter, a sgRNA controlled constitutively from PpflB S. aureus promoter to clone a guide between two BsaI sites and oriT on pLZ12 vector pLZ12
	pLY9	dcas9 (Other), tetR (Other), pspFΔHTH::λN22plus (Synthetic), sfgfp (Synthetic)	Ptet, Ptet, Anderson promoter: J23106, PpspA-2G6	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	A CRISPR activation device with the necessary genes (dcas9 controlled by Ptet, pspFΔHTH::λN22plus controlled by promoter J23106), and the reporter part (PpspA-2G6 with sfgfp::ASV). pSB4A3mut
	pLY53	dcas9 (Other), tetR (Other), pspFΔHTH::λN22plus (Synthetic), sfgfp (Synthetic)	Ptet, Ptet, Anderson promoter: J23106, PpspA-LEA1B1	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	A CRISPR activation device with the necessary genes (dcas9 controlled by Ptet, pspFΔHTH::λN22plus controlled by promoter J23106), and the reporter part (PpspA-LEA1B1 with sfgfp::ASV). pSB4A3mut
	pLY54	dcas9 (Other), tetR (Other), pspFΔHTH::λN22plus (Synthetic), sfgfp (Synthetic)	Ptet, Ptet, Anderson promoter: J23106, PpspA-LEA2B2	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	A CRISPR activation device with the necessary genes (dcas9 controlled by Ptet, pspFΔHTH::λN22plus controlled by promoter J23106), and the reporter part (PpspA-LEA2B2 with sfgfp::ASV). pSB4A3mut
	pLY55	dcas9 (Other), tetR (Other), pspFΔHTH::λN22plus (Synthetic), sfgfp (Synthetic)	Ptet, Ptet, Anderson promoter: J23106, PpspA-LEA3B3	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	A CRISPR activation device with the necessary genes (dcas9 controlled by Ptet, pspFΔHTH::λN22plus controlled by promoter J23106), and the reporter part (PpspA-LEA3B3 with sfgfp::ASV). pSB4A3mut
	pLY56	dcas9 (Other), tetR (Other), pspFΔHTH::λN22plus (Synthetic), sfgfp (Synthetic)	Ptet, Ptet, Anderson promoter: J23106, PpspA-LEA4B4	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	A CRISPR activation device with the necessary genes (dcas9 controlled by Ptet, pspFΔHTH::λN22plus controlled by promoter J23106), and the reporter part (PpspA-LEA4B4 with sfgfp::ASV). pSB4A3mut
	pLY57	dcas9 (Other), tetR (Other), pspFΔHTH::λN22plus (Synthetic), sfgfp (Synthetic)	Ptet, Ptet, Anderson promoter: J23106, PpspA-LEA5B5	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	A CRISPR activation device with the necessary genes (dcas9 controlled by Ptet, pspFΔHTH::λN22plus controlled by promoter J23106), and the reporter part (PpspA-LEA5B5 with sfgfp::ASV). pSB4A3mut
	pLY59	dcas9 (Synthetic), tetR (Other), sfgfp (Synthetic), pspFΔHTH::MCP (Other)	Ptet, Ptet, PpspA-2G6, Anderson promoter: J23106	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	A CRISPR activation device with the necessary genes (dcas9 controlled by Ptet, pspFΔHTH::MCP controlled by promoter J23106), and the reporter part (PpspA-2G6 with sfgfp::ASV). pSB4A3mut
	pLY61	dxcas9 (3.7) (Synthetic), tetR (Other), pspFΔHTH::λN22plus (Synthetic), sfgfp (Synthetic)	Ptet, Ptet, Anderson promoter: J23106, PpspA-LEA1B1	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	A CRISPR activation device with the necessary genes (dxcas9 3.7 controlled by Ptet, pspFΔHTH::λN22plus controlled by promoter J23106), and the reporter part (PpspA-LEA1B1 with sfgfp::ASV). pSB4A3mut
	pLY70	dxcas9 (3.7) (Other), tetR (Other), rhaS (Other), pspFΔHTH::λN22plus (Synthetic), sfgfp (Synthetic)	Ptet, Ptet, Pcon, PrhaB, PpspA-LEA3B3	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	A CRISPR activation device with the necessary genes (dxcas9 controlled by Ptet, pspFΔHTH::λN22plus controlled by promoter PrhaB), and the reporter part (PpspA-LEA3B3 with sfgfp::ASV). pSB4A3mut
	pLY152	dcas9 (Other), tetR (Other), sfgfp (Synthetic)	Ptet, Ptet, PpspA-2G6	Wang	Engineered CRISPRa enables programmable eukaryote-like gene activation in bacteria Nature Communications 10, August 2019	dcas9 generator and reporter circuit (PpspA-2G6 with sfgfp::ASV) pSB4A3mut
	pCD442	dCas9, MCP-SoxS(R93A/S101A) (Other)		Zalatan	Effective CRISPRa-mediated control of gene expression in bacteria must overcome strict target site requirements. Nat Commun. 2020 Apr 1;11(1):1618. doi: 10.1038/s41467-020-15454-y.	dCas9, MCP-SoxS(R93A/S101A) p15A vector
	pCK005.6	dCas9, MCP-SoxS(R93A/S101A), J106 scRNA (Other)		Zalatan	Effective CRISPRa-mediated control of gene expression in bacteria must overcome strict target site requirements. Nat Commun. 2020 Apr 1;11(1):1618. doi: 10.1038/s41467-020-15454-y.	dCas9, MCP-SoxS(R93A/S101A), J106 scRNA p15A vector
	pdCas9-AsiA	dCas9-AsiA (Synthetic)	TetO	Wang	Programmable CRISPR-Cas transcriptional activation in bacteria (unpublished)	Expressing CasTA of dCas9 fused AsiA pdCas9-bacteria
	pdCas9- AsiA_m1.1	dCas9-AsiA_m1.1 (Synthetic)	TetO	Wang	Programmable CRISPR-Cas transcriptional activation in bacteria (unpublished)	Expressing evolved CasTA of dCas9 fused AsiA variant 1.1 pdCas9-bacteria
	pdCas9- AsiA_m2.1	dCas9-AsiA_m2.1 (Synthetic)	TetO	Wang	Programmable CRISPR-Cas transcriptional activation in bacteria (unpublished)	Expressing evolved CasTA of dCas9 fused AsiA variant 2.1 pdCas9-bacteria

Interfere

Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down gene expression by interfering with transcription. Design your gRNA to target your gene of interest’s promoter/enhancer or the beginning of the coding sequence. If the plasmid you’re using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-repressor to your specific locus.

	Plasmid	Gene/Insert	Promoter	PI	Publication	Hidden Extra Search Info
	pMJ841	Cas9 (Other)	T7	Doudna	A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012 Jun 28.	pEC-K-MBP Tags / Fusion Proteins MBP (N terminal on backbone) His6 (N terminal on backbone)
	pdCas9-bacteria	dCas9 (bacteria) (Other)	pLtetO-1	Qi	Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022.	aTc-inducible expression of a catalytically inactive bacterial Cas9 (S. pyogenes) for bacterial gene knockdown p15A vector
	pdCas9	tracrRNA (Other), dcas9 (Other), CRISPR array		Marraffini	Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic Acids Res. 2013 Jun 12.	Expresses the tracrRNA, the dCas9 catalytic site mutant and a CRISPR array designed for the easy cloning of new spacers. pACYC184
	10xHis-MBP-TEV-S. pyogenes dCas9 M1C D10A C80S H840A C574S	dCas9 M1C D10A C80S H840A C574S (Other)		Doudna	Programmable RNA recognition and cleavage by CRISPR/Cas9. Nature. 2014 Sep 28. doi: 10.1038/nature13769.	dCas9 with single cysteine residue (M1C) for site-specific labelling pHMGWA
	pAN-PTet-dCas9	dCas9 (Other)	Ptet	Voigt	Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks. Mol Syst Biol. 2014 Nov 24;10:763. doi: 10.15252/msb.20145735.	controls the expression of S. pyogenes dCas9 from a Tc‐inducible PTet promoter. unknown
	pCRISPathBrick		Constitutive native promoters	Koffas	CRISPathBrick: Modular Combinatorial Assembly of Type II-A CRISPR Arrays for dCas9-Mediated Multiplex Transcriptional Repression in E. coli. ACS Synth Biol. 2015 Mar 30.	E. coli vector for expression of S. pyogenes dCas9, tracrRNA, and nontargeting CRISPR array with BsaI site for inserting user-defined spacer-repeat bricks pdCas9-Marraffini (pACYC184)
	MSP712	mammalian codon-optimized Streptococcus pyogenes dCas9 (D10A/H840A)-NLS-3XFlag, and SpCas9 gRNA (Other)	T7 (x2)	Joung	Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. 2015 Jun 22. doi: 10.1038/nature14592.	Bacterial expression plasmid for Sp-dCas9 & sgRNA (need to clone in spacer into BsaI sites): T7-humanSpdCas9(D10A/H840A)-T7-BsaIcassette-Sp-sgRNA pACYCDuet-1 Tags / Fusion Proteins NLS (C terminal on insert) 3x FLAG (C terminal on insert)
	pMM704	dCas9, LacI, sgRNA		Lu	Programming a Human Commensal Bacterium, Bacteroides thetaiotaomicron, to Sense and Respond to Stimuli in the Murine Gut Microbiota. Cell Syst. 2015 Jul 29;1(1):62-71. doi: 10.1016/j.cels.2015.06.001.	IPTG-inducible CRISPRi vector targeting BT1854, pNBU2 backbone, AmpR pExchange-tdk
	pMD19T-psba1-Ppsba2-dCas9-SpR	dCas9 from S. pyogenes (Other)	PpsbA2	Hudson	Multiple Gene Repression in Cyanobacteria Using CRISPRi. ACS Synth Biol. 2015 Dec 28.	Contains dCas9 from S. pyogenes under constitutive promoter. Suicide vector inserts into psba1 site of Synechocystis. Carries spectinomycin resist. Recommend E. coli Copy cutter for propogation. pMD19T simple
	pMD19T-psba1-TetR-PL22-dCas9-SpR	dCas9 from S. pyogenes (Other)	PL22	Hudson	Multiple Gene Repression in Cyanobacteria Using CRISPRi. ACS Synth Biol. 2015 Dec 28.	Contains dCas9 from S. pyogenes under aTc inducible promoter. Suicide vector inserts into psba1 site of Synechocystis. Carries spectinomycin resist. Recommend E. coli Copy cutter for propogation. pMD19T simple
	pdCas9-M-C4	Repressor C4 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant C4. pdCas9
	pdCas9-M-3F2	Repressor C4 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 3F2. pdCas9
	pdCas9-M-3H5	Repressor 3H5 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 3H5. pdCas9
	pdCas9-M-1B6	Repressor 1B6 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 1B6. pdCas9
	pdCas9-M-4F2	Repressor 4F2 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 4F2. pdCas9
	pdCas9-M-5F5	Repressor 5F5 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 5F5. pdCas9
	pdCas9-M-1D4	Repressor 1D4 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 1D4. pdCas9
	pdCas9-M-4A6	Repressor 4A6 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 4A6. pdCas9
	pdCas9-M-3A2	Repressor 3A2 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 3A2. pdCas9
	pdCas9-M-1E4	Repressor 1E4 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant 1E4. pdCas9
	pdCas9-M-G6	Repressor G6 (orthogonal T7-lac repressor) (Synthetic)	Constitutive wild-type S. pyogenes promoter	Koffas	Rapid generation of CRISPR/dCas9-regulated, orthogonally repressible hybrid T7-lac promoters for modular, tuneable control of metabolic pathway fluxes in Escherichia coli. Nucleic Acids Res. 2016 Apr 13. pii: gkw231.	CRISPR synthetic transcription factor repressor plasmid encoding dCas9, tracrRNA, and a single spacer CRISPR array encoding crRNA for orthogonal repression of T7-lac promoter variant G6. pdCas9
	pZ8-T_dCas9	dcas9	ptac	Lu	Corynebacterium glutamicum Metabolic Engineering with CRISPR Interference (CRISPRi). ACS Synth Biol. 2016 Feb 16.	pZ8-1 plasmid carrying dcas9, driven by the IPTG-inducible Ptac promoter, KanR pZ8-1
	pZ8-P_dCas9	dcas9	Propionate inducible promoter (prp)	Lu	Corynebacterium glutamicum Metabolic Engineering with CRISPR Interference (CRISPRi). ACS Synth Biol. 2016 Feb 16.	pZ8-1 plasmid carrying dcas9 driven by the propionate-inducible prpD2 promoter (PprpD2), KanR pZ8-Prp
	pJMP1	dCas9 (Other)	xylA	Gross	A Comprehensive, CRISPR-based Functional Analysis of Essential Genes in Bacteria. Cell. 2016 Jun 2;165(6):1493-506. doi: 10.1016/j.cell.2016.05.003. Epub 2016 May 26.	Bacillus subtilis dCas9 expression vector; integrates into lacA/ganA pAX01
	pCas2A	dCas9	PEZ3	Pfleger	CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007.	dCas9 expressed from aTc-inducible promoter in acsA, A RBS: AGGAGA pACSA
	pCas2B	dCas9	PEZ3	Pfleger	CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007.	dCas9 expressed from aTc-inducible promoter in acsA, B RBS: TCGAGA pACSA
	pCas2C	dCas9	PEZ3	Pfleger	CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007.	dCas9 expressed from aTc-inducible promoter in acsA, C RBS: TGGACA pACSA
	pCas2D	dCas9	PEZ3	Pfleger	CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007.	dCas9 expressed from aTc-inducible promoter in acsA, D RBS: AGGACG pACSA
	pCas2E	dCas9	PEZ3	Pfleger	CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007.	dCas9 expressed from aTc-inducible promoter in acsA, E RBS: AGGGCG pACSA
	pCas2F	dCas9	PEZ3	Pfleger	CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007.	dCas9 expressed from aTc-inducible promoter in acsA, F RBS: TGGGCG pACSA
	pCas7	dCas9	c225	Pfleger	CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007.	dCas9 expressed from low constituve promoter, c225, in acsA pACSA
	pCas8	dCas9	none	Pfleger	CRISPR interference as a titratable, trans-acting regulatory tool for metabolic engineering in the cyanobacterium Synechococcus sp. strain PCC 7002. Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 10.1016/j.ymben.2016.07.007.	dCas9 without a promoter in acsA pACSA
	pRH2502	dcas9 (Other)	uv15tetO	Husson	Investigating essential gene function in Mycobacterium tuberculosis using an efficient CRISPR interference system. Nucleic Acids Res. 2016 Jul 12. pii: gkw625.	Expression of dcas9 D10A H840A from a TetR-regulated uvtetO promoter pTC-0X-1L
	pAW019-2	dcas9 (Other)	PxylA (B. megaterium)	Chou	Development of a CRISPR-Cas9 toolkit for comprehensive engineering of Bacillus subtilis. Appl Environ Microbiol. 2016 Jun 3. pii: AEM.01159-16.	Inducible dCas9 integration vector for Bacillus subtilis pAX01 (ColE1)
	pUC18-mini-Tn7T-Lac-dCas9 (Ptac)	S. pasteurianus dCas9 (Other)		Prather	A Robust CRISPR Interference Gene Repression System in Pseudomonas. J Bacteriol. 2018 Mar 12;200(7). pii: JB.00575-17. doi: 10.1128/JB.00575-17. Print 2018 Apr 1.	Tn7 integrating plasmid with S. pasteurianus dCas9 expressed from the Ptac promoter for expression in P. putida and P. fluorescens pUC18
	pUC18-mini-Tn7T-Plac-dCas9 (Plac)	S. pasteurianus dCas9 (Other)		Prather	A Robust CRISPR Interference Gene Repression System in Pseudomonas. J Bacteriol. 2018 Mar 12;200(7). pii: JB.00575-17. doi: 10.1128/JB.00575-17. Print 2018 Apr 1.	Tn7 integrating plasmid with S. pasteurianus dCas9 expressed from the Plac promoter for expression in P. aeruginosa pUC18
	Biomolecular FeedBack (FB)	pJ23100mut-dCas9-T- phtpG1-sgRNA (Other)		Ellis	Burden-driven feedback control of gene expression. Nat Methods. 2018 Mar 26. pii: nmeth.4635. doi: 10.1038/nmeth.4635.	This construct encodes for two units. The first contains a constitutively expressed dCas9 protein. The second contains an sgRNA cassette under the control of the burden-responsive htpG1 promoter. pAZ16 ( Plasmid has got AmpR and p15A origin)
	pSET-dCas9	dCas9 (Synthetic)	ermE*p	Lu	CRISPR/dCas9-Mediated Multiplex Gene Repression in Streptomyces. Biotechnol J. 2018 Jun 3. doi: 10.1002/biot.201800121.	Expression of the dCas9 gene in Streptomyces pSET152
	pSET-dCas9-actII-4-NT-S1	dCas9 (Synthetic), sgRNA	ermE*p, J23119	Lu	CRISPR/dCas9-Mediated Multiplex Gene Repression in Streptomyces. Biotechnol J. 2018 Jun 3. doi: 10.1002/biot.201800121.	Repression of gene expression in Streptomyces by CRISPRi pSET152
	pCT310	dCas9 (Other)	T7	Tjian	dCas9-targeted locus-specific protein isolation method identifies histone gene regulators. Proc Natl Acad Sci U S A. 2018 Mar 20;115(12):E2734-E2741. doi: 10.1073/pnas.1718844115. Epub 2018 Mar 5.	Bacterial expression of 6His-dCas9-3XFlag pET302/NT-His
	pdCas9-J23111	dCas9 (Other)		Zhang	Pooled CRISPR interference screening enables genome-scale functional genomics study in bacteria with superior performance. Nat Commun. 2018 Jun 26;9(1):2475. doi: 10.1038/s41467-018-04899-x.	Plasmid constitutively expressing dCas9 protein N.A.
	pdCas9-J23109	dCas9 (Other)		Zhang	Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572.	Plasmid constitutively expressing dCas9 protein N.A.
	peSpdCas9-J23109	eSpdCas9 (Other)		Zhang	Improved sgRNA design in bacteria via genome-wide activity profiling. Nucleic Acids Res. 2018 Aug 21;46(14):7052-7069. doi: 10.1093/nar/gky572.	Plasmid constitutively expressing eSpdCas9 protein N.A.
	Native promoter dCas9	Native promoter and dCas9 (Other)	S. pyogenes Cas9 native promoter	Barrick	Synthetic Genome Defenses against Selfish DNA Elements Stabilize Engineered Bacteria against Evolutionary Failure. ACS Synth Biol. 2019 Mar 15;8(3):521-531. doi: 10.1021/acssynbio.8b00426. Epub 2019 Feb 15.	dCas9 expression unit plasmid. The dCas9 expression unit plasmids contain connector ConL1, ConRE, and one dCas9 transcriptional unit. pYTK095
	TDK promoter dCas9	TDK gene promoter and dCas9 (Other)	TDK gene promoter	Barrick	Synthetic Genome Defenses against Selfish DNA Elements Stabilize Engineered Bacteria against Evolutionary Failure. ACS Synth Biol. 2019 Mar 15;8(3):521-531. doi: 10.1021/acssynbio.8b00426. Epub 2019 Feb 15.	dCas9 expression unit plasmid. The dCas9 expression unit plasmids contain connector ConL1, ConRE, and one dCas9 transcriptional unit. pYTK095
	pNS38-SadCas9-SNAP	Cas9 (Other)		Schwank	Covalent linkage of the DNA repair template to the CRISPR-Cas9 nuclease enhances homology-directed repair. Elife. 2018 May 29;7. pii: 33761. doi: 10.7554/eLife.33761.	Bacterial vector for expression of Snap-tagged Staphylococcus aureus dCas9 pET His6 MBP N10 TEV LIC cloning vector (2C-T)
	PLJR962	Sth1 sgRNA scaffold (Other), Sth1 dCas9 (Other), TetR (Other), L5 attP (Other), L5 Int (Other), KanR (Other)		Fortune	Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform. Nat Microbiol. 2017 Feb 6;2:16274. doi: 10.1038/nmicrobiol.2016.274.	Sth1 dCas9 TetR and KanR L5 Int attP for M. smegmatis custom design
	PLJR965	Sth1 sgRNA scaffold (Other), Sth1 dCas9 (Other), TetR (deleted TetON) MtbCO (Other), L5 attP (Other), L5 Int (Other), KanR (Other)		Fortune	Programmable transcriptional repression in mycobacteria using an orthogonal CRISPR interference platform. Nat Microbiol. 2017 Feb 6;2:16274. doi: 10.1038/nmicrobiol.2016.274.	Sth1 dCas9 TetR and KanR L5 Int attP for M. tuberculosis custom design
	FR-E01	none		Bikard	Genome-wide CRISPR-dCas9 screens in E. coli identify essential genes and phage host factors. PLoS Genet. 2018 Nov 7;14(11):e1007749. doi: 10.1371/journal.pgen.1007749. eCollection 2018 Nov.	Expression of dCas9 gene under the control of a Ptet promoter in the E. coli chromosome none
	p15A_J23108_SpydCas9_CmR	dCas9 from S. pyogenes		Noireaux	An educational module to explore CRISPR technologies with a cell-free transcription-translation system (unpublished)	Expresses S. pyogenes dCas9 pBAD33
	pJMP1161	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	rfp "test" strain R6Kgamma
	pJMP1171	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	rfp "test" strain R6Kgamma
	pJMP1185	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	rfp "test" strain R6Kgamma
	pJMP1189	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	rfp "test" strain R6Kgamma
	pJMP1219	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	rfp "test" strain R6Kgamma
	pJMP1223	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	rfp "test" strain R6Kgamma
	pJMP1237	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	for cloning new sgRNAs R6Kgamma
	pJMP1335	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	rfp "test" strain R6Kgamma
	pJMP1337	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	for cloning new sgRNAs R6Kgamma
	pJMP1339	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	for cloning new sgRNAs R6Kgamma
	pJMP1354	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	for cloning new sgRNAs R6Kgamma
	pJMP1356	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	for cloning new sgRNAs R6Kgamma
	pJMP1358	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	for cloning new sgRNAs R6Kgamma
	pJMP1360	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	for cloning new sgRNAs R6Kgamma
	pJMP1363	dcas9 (Other)		Gross	Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 10.1038/s41564-018-0327-z.	for stabilizing ICE in the presence of rapI expression R6Kgamma
	pC0.017	dCas9 (Other)		McCormick	CyanoGate: A Modular Cloning Suite for Engineering Cyanobacteria Based on the Plant MoClo Syntax. Plant Physiol. 2019 May;180(1):39-55. doi: 10.1104/pp.18.01401. Epub 2019 Feb 28.	Level 0 Part. CDS pICH41308
	pC0.018	dCas9+deg-tag (Other)		McCormick	CyanoGate: A Modular Cloning Suite for Engineering Cyanobacteria Based on the Plant MoClo Syntax. Plant Physiol. 2019 May;180(1):39-55. doi: 10.1104/pp.18.01401. Epub 2019 Feb 28.	Level 0 Part. CDS pICH41308
	C41m	dCas9 (Synthetic)		Murray	CIDAR MoClo Extension (unpublished)	MoClo golden gate assembly CD part for dCas9 (Catalytically-inactive S. pyogenes Cas9, for gRNA-targeted repression). Please see Supplemental Documents for annotated Genbank file. DVA
	C114m	dCas9 with no bbsI or BsaI sites (Synthetic)		Murray	CIDAR MoClo Extension (unpublished)	MoClo golden gate assembly CD part for Cas9 (S. pyogenes gRNA-targeted endonuclease Cas9, with bbsI cut sites removed synonymously). Please see Supplemental Documents for annotated Genbank file. DVA
	pD2-dcas9-J23101-GFP	dCas9 (Synthetic)		Poh	Regulating exopolysaccharide gene wcaF allows control of Escherichia coli biofilm formation. Sci Rep. 2018 Sep 3;8(1):13127. doi: 10.1038/s41598-018-31161-7.	Constitutive dCas9 and GFP expression plasmid pBbA2c
	pFD116	Ptet (Other), dcas9 (Other), PpflB sgRNA BsaI (Synthetic)		Bikard	Gene silencing with CRISPRi in bacteria and optimization of dCas9 expression levels. Methods. 2019 Aug 1. pii: S1046-2023(18)30497-3. doi: 10.1016/j.ymeth.2019.07.024.	pFD116 carries dcas9 controlled by an aTc-inducible Ptet promoter, a sgRNA controlled constitutively from PpflB S. aureus promoter to clone a guide between two BsaI sites and oriT on pLZ12 vector pLZ12
	pCRISPR-dCas9	streptomyces codon optimized spdCas9, sgRNA casette (Other)	ermE*/tipA	Weber	CRISPR-Cas9 Based Engineering of Actinomycetal Genomes. ACS Synth Biol. 2015 Sep 18;4(9):1020-9. doi: 10.1021/acssynbio.5b00038. Epub 2015 Apr 7.	Gene knockdown for actinomycetes pGM1190
	pJUMP29[dCas9]-1A(lacZ)			French	Joint Universal Modular Plasmids (JUMP): A flexible and comprehensive platform for synthetic biology bioRxiv, October 10, 2019	Level 1 vector. With lacZ as alternative cloning reporter.OriV 9 (pBBR322/ROP; medium copy number). Constitutive dCas9 in downstream secondary site. pJUMP29-1A
	pJUMP18-dCas9_O	Part dCas9_O (Synthetic)		French	Joint Universal Modular Plasmids (JUMP): A flexible and comprehensive platform for synthetic biology bioRxiv, October 10, 2019	Basic Part O- ORF; Catalytically dead mutant of the Cas9 from Streptococcus pyogenes. pJUMP18-Uac
	pAH-CTX1-rhadCas9	Burkholderia cenocepacia codon-optimized dCas9 (Synthetic)	PrhaBAD	Cardona	A broad-host-range CRISPRi toolkit for silencing gene expression in Burkholderia. ACS Synth Biol. 2019 Sep 6. doi: 10.1021/acssynbio.9b00232.	Derived from pAH-CTX1-rha. The Burkholderia cenocepacia codon-optimized dCas9 cloned downstream of PrhaBAD. Hence, dCas9 is controlled by the rhamnose-inducible promoter system. pAH-CTX1-rha
	pAH-CTX1-rhadCas9-native	S. pyogenes dCas9 (Other)	PrhaBAD	Cardona	A broad-host-range CRISPRi toolkit for silencing gene expression in Burkholderia. ACS Synth Biol. 2019 Sep 6. doi: 10.1021/acssynbio.9b00232.	Derived from pAH-CTX1-rha. The native Streptococcus pyogenes dCas9 gene cloned downstream of PrhaBAD. Non-codon-optimized dCas9 version of pAH-CTX1-rhadCas9. pAH-CTX1-rha
	pXGFPC-5 Pxyl-dcas9 (Spy)	dcas9 (Streptococcus pyogenes) (Other)	xylose	Laub	A CRISPR Interference System for Efficient and Rapid Gene Knockdown in Caulobacter crescentus mBio	expresses dcas9 from Streptococcus pyogenes; repressed with 0.2% glucose, induced with 0.3% xylose; for homologous recombination at the xylose locus in Caulobacter crescentus; tetracycline resistance pXGFPC-5
	pXGFPC-5 Pxyl-dcas9 (Sth3)	dcas9 (Streptococcus thermophilus #3) (Other)	xylose	Laub	A CRISPR Interference System for Efficient and Rapid Gene Knockdown in Caulobacter crescentus mBio	expresses dcas9 (Streptococcus thermophilus #3); repressed with 0.2% glucose, induced with 0.3% xylose; for homologous recombination at the xyl locus in Caulobacter crescentus; tetracycline resistance pXGFPC-5
	pXGFPC-5 Pxyl-dcas9 (Spa)	dcas9 (Streptococcus pasteurianus) (Other)	xylose	Laub	A CRISPR Interference System for Efficient and Rapid Gene Knockdown in Caulobacter crescentus mBio	expresses dcas9 (Streptococcus pasteurianus); repressed with 0.2% glucose, induced with 0.3% xylose; for homologous recombination at the xylose locus in Caulobacter crescentus; tetracycline resistance pXGFPC-5
	pVCERC-5 Pvan-dcas9 (Sth3)	dcas9 (Streptococcus thermophilus #3) (Other)	vanillate	Laub	A CRISPR Interference System for Efficient and Rapid Gene Knockdown in Caulobacter crescentus mBio	expresses dcas9 from Streptococcus thermophilus #3; induced with vanillate; for homologous recombination at the vanillate locus in Caulobacter crescentus; tetracycline resistance pVCERC-5
	pJ1996_v2	dCas9 & Csy4		Schaerli	Multistable and dynamic CRISPRi-based synthetic circuits. Nat Commun. 2020 Jun 2;11(1):2746. doi: 10.1038/s41467-020-16574-1.	dCas9 & Csy4 pCDF
	pJ2018	dCas9, LuxR & Csy4		Schaerli	Multistable and dynamic CRISPRi-based synthetic circuits. Nat Commun. 2020 Jun 2;11(1):2746. doi: 10.1038/s41467-020-16574-1.	dCas9, LuxR & Csy4 pCDF
	pMSP3545-dCas9Str	dCas9Str (Other)	nisA	Kline	Multiplex CRISPRi-Cas9 silencing of planktonic and stage-specific biofilm genes in Enterococcus faecalis (unpublished)	Expresses dCas9str under nisin-inducible nisA promoter in pMSP3545 backbone pMSP3545
	pSM-dCas9::Ap			Ng	CRISPRi-mediated Programming Essential Gene can as a Direct Enzymatic Performance Evaluation & Determination (DEPEND) System. Biotechnol Bioeng. 2020 May 27. doi: 10.1002/bit.27443.	Shuttle vector for expression dCas9, mobilization cluster, expresses the tracrRNA and a CRISPR array designed for the easy cloning of new spacers. pSM-dCas9

RNA Targeting

Type VI CRISPR systems, including the enzymes Cas13a/C2c2 and Cas13b, target RNA rather than DNA. In bacteria, once they have recognized and cleaved the target RNA sequence, they adopt an enzymatically active state and can bind and cleave additional RNAs regardless of homology to the crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which require that each DNA target have high sequence identity to the spacer sequence and contain a PAM sequence just downstream of the sequence to be cleaved. This non-specific cleavage is thought to activate programmed cell death or dormancy for phage-infected bacterial cells so as to limit the spread of infection throughout the entire population.

	Plasmid	Promoter	PI	Publication	Hidden Extra Search Info
	pZ003 (LshC2c2 locus)		Zhang	Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems. Mol Cell. 2015 Nov 5;60(3):385-97. doi: 10.1016/j.molcel.2015.10.008. Epub 2015 Oct 22.	Expresses LshC2c2 full locus pACYC184
	pZ004 (LseC2c2 locus)		Zhang	Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems. Mol Cell. 2015 Nov 5;60(3):385-97. doi: 10.1016/j.molcel.2015.10.008. Epub 2015 Oct 22.	Expresses LseC2c2 locus with one spacer in array pET28a
	pC001 - huLshC2C2-MBP for bacterial expression	T7	Zhang	C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Jun 2. pii: aaf5573.	Expresses human codon-optimized LshC2c2 for purification in E. coli BPV00356 pET21GG2-His(6)-MBP_Flag-Avi
	pC002 - LshC2C2 locus into pACYC184		Zhang	C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Jun 2. pii: aaf5573.	Endogenous LshC2c2 locus cloned into pACYC184 pACYC184
	pC003 - LshC2C2 locus into pACYC184 for spacer cloning		Zhang	C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Science. 2016 Jun 2. pii: aaf5573.	LshC2c2 locus in pACYC184 with BsaI sites for spacer cloning pACYC184
	p2CT-His-MBP-Lbu_C2c2_WT		Doudna	Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.	Bacterial expression of WT L. buccalis C2c2 CRISPR effector. p2CT
	p2CT-His-MBP-Lbu_C2c2_R472A_H477A		Doudna	Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.	Bacterial expression of L. buccalis C2c2 CRISPR effector (HEPN nuclease 1 inactive mutant). p2CT
	p2CT-His-MBP-Lbu_C2c2_R1048A_H1053A		Doudna	Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.	Bacterial expression of L. buccalis C2c2 CRISPR effector (HEPN nuclease 2 inactive mutant). p2CT
	p2CT-His-MBP-Lbu_C2c2_R472A_H477A_R1048A_H1053A		Doudna	Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.	Bacterial expression of L. buccalis C2c2 CRISPR effector (double HEPN nuclease 1 and 2 inactive mutant). p2CT
	p2CT-His-MBP-Lse_C2c2_WT		Doudna	Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.	Bacterial expression of WT L. seeligeri C2c2 CRISPR effector p2CT
	p2CT-His-MBP-Lsh_C2c2_WT		Doudna	Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.	Bacterial expression of WT L. shahii C2c2 CRISPR effector p2CT
	p2CT-His-MBP-Lbu_C2c2_R1079A		Doudna	Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection. Nature. 2016 Sep 26. doi: 10.1038/nature19802.	Bacterial expression of L. buccalis C2c2 CRISPR effector (R1079A- pre-crRNA processing mutant) p2CT
	pBZCas13b	Lac	Zhang	Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5.	Bacterial expression for bzCas13b, driven by the lac promoter, and DR-spacer-DR sequence driven by js23119. New spacer sequences can be cloned in between the DRs by digesting the plasmid with BsaI. pACYC184
	pBZCas13b-HEPN	Lac	Zhang	Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5.	Bacterial expression for bzCas13b and crRNA with both HEPN domains mutated. New spacers can be cloned by digesting with BsaI. pACYC184
	pPbcas13b	Lac	Zhang	Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase Differentially Regulated by Accessory Proteins Csx27 and Csx28. Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5.	Bacterial expression for pbCas13b, driven by the lac promoter, and DR-spacer-DR sequence driven by js23119. New spacer sequences can be cloned in between the DRs by digesting the plasmid with BsaI. pACYC184
	pC013 - Twinstrep-SUMO-huLwCas13a		Zhang	Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.	Twinstrep-SUMO-huLwCas13a for recombinant protein bacterial expression. Insert is human codon optimized but expresses well in bacteria. pET
	p2CT-His-MBP-Lbu_C2c2_E299A		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lbu_C2c2_K310A		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lbu_C2c2_N314A		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lbu_C2c2_R1072A		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lbu_C2c2_D1078A		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lbu_C2c2_K1080A		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lbu_C2c2_K1082A		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lbu_C2c2_K1087A		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lwa_Cas13a_WT		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lne_Cas13a_WT		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Lba_Cas13a_WT		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Ere_Cas13a_WT		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Cam_Cas13a_WT		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Rca_Cas13a_WT		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Hhe_Cas13a_WT		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	p2CT-His-MBP-Ppr_Cas13a_WT		Doudna	RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008.	Bacterial expression for Cas13a p2CT Tag / Fusion Protein His6-MBP-TEV site (N terminal on backbone)
	pC009 LshCas13a locus with targeting spacer		Zhang	Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.	LshCas13a locus into pACYC184 with targeting spacer pACYC184
	pC010 LshCas13a locus with nontargeting spacer		Zhang	Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.	LshCas13a locus into pACYC184 with nontargeting spacer pACYC184
	pC011 LwCas13a locus with targeting spacer		Zhang	Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.	LwCas13a locus into pACYC184 with targeting spacer pACYC184
	pC012 LwCas13a locus with nontargeting spacer		Zhang	Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.	LwCas13a locus into pACYC184 with nontargeting spacer pACYC184
	pC018 - LshCas13a from Leptotrichia shahii		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses LshCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC019 - LwCas13a from Leptotrichia wadei		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses LwCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC020 - LseCas13a from Listeria seeligeri		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses LseCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC021 - LbmCas13a from Lachnospiraceae bacterium MA2020		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses LbmCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC022 - LbnCas13a from Lachnospiraceae bacterium NK4A179		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses LbnCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC023 - CaCas13a from [Clostridium] aminophilum DSM 10710		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses CaCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC024 - CgCas13a from Carnobacterium gallinarum DSM 4847		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses CgCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC025 - Cg2Cas13a from Carnobacterium gallinarum DSM 4847		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses Cg2Cas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC026 - PpCas13a from Paludibacter propionicigenes WB4		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses PpCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC027 - LweCas13a from Listeria weihenstephanensis FSL R9-0317		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses LweCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC028 - LbfCas13a from Listeriaceae bacterium FSL M6-0635		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses LbfCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC029 - Lw2Cas13a from Leptotrichia wadei F0279		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses Lw2Cas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC030 - RcsCas13a from Rhodobacter capsulatus SB 1003		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses RcsCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC031 - RcrCas13a from Rhodobacter capsulatus R121		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses RcrCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pC032 - RcdCas13a from Rhodobacter capsulatus DE442		Zhang	RNA targeting with CRISPR-Cas13. Nature. 2017 Oct 4. doi: 10.1038/nature24049.	Expresses RcdCas13a for bacterial expression and contains backbone for spacer cloning pACYC184
	pDuBir-Lbu-dCas13a-avitag	T7	O'Connell	RNA Binding and HEPN-Nuclease Activation Are Decoupled in CRISPR-Cas13a. Cell Rep. 2018 Jul 24;24(4):1025-1036. doi: 10.1016/j.celrep.2018.06.105.	Dual expression of Lbu-dCas13a with a C-terminal avi-tag and BirA to biotinylate Lbu-dCas13a pDuBir
	pET28a-MH6-EsCas13d	lac	Biotechnologies	Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein. Mol Cell. 2018 Mar 9. pii: S1097-2765(18)30173-4. doi: 10.1016/j.molcel.2018.02.028.	Expresses E. coli codon optimized EsCas13d in the pET28a backbone. pET-28a(+)
	pET28a-MH6-RspCas13d_RspCasWYL1	lac	Biotechnologies	Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein. Mol Cell. 2018 Mar 9. pii: S1097-2765(18)30173-4. doi: 10.1016/j.molcel.2018.02.028.	Expresses E. coli codon optimized RspCas13d and RspCasWYL1 in the pET28a backbone. pET-28a(+)
	pET28a-MH6-RspCas13d	lac	Biotechnologies	Cas13d Is a Compact RNA-Targeting Type VI CRISPR Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein. Mol Cell. 2018 Mar 9. pii: S1097-2765(18)30173-4. doi: 10.1016/j.molcel.2018.02.028.	Expresses E. coli codon optimized RspCas13d in the pET28a backbone. pET-28a(+)
	pC0057 BzoCas13 (B01) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses BzoCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0058 PinCas13 (B02) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses PinCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0059 PbuCas13 (B03) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses PbuCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0060 AspCas13 (B04) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses AspCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0061 PsmCas13 (B05) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses PsmCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0062 RanCas13 (B06) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses RanCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0063 PauCas13 (B07) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses PauCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0065 Pin2Cas13 (B09) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses Pin2Cas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0067 PguCas13 (B11) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses PguCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0068 PspCas13 (B12) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses PspCas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0070 Pin3Cas13 (B15) His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses Pin3Cas13b from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pC0072 LbuCas13a His6-TwinStrep-SUMO-BsaI		Zhang	Multiplexed and portable nucleic acid detection platform with Cas13, Cas12a, and Csm6. Science. 2018 Apr 27;360(6387):439-444. doi: 10.1126/science.aaq0179. Epub 2018 Feb 15.	Expresses LbuCas13a from a SUMO-tagged bacterial expression construct SUMO bacterial expression vector
	pLsCas13aGG		Beisel	Modular one-pot assembly of CRISPR arrays enables library generation and reveals factors influencing crRNA biogenesis. Nat Commun. 2019 Jul 3;10(1):2948. doi: 10.1038/s41467-019-10747-3.	Backbone plasmid for generating CRISPR arrays for LsCas13a. Contains a GFP-dropout cassette and a direct repeat. pBAD18
	pET-28b-RfxCas13d-His	T7	Moreno-Mateos	CRISPR-Cas13d Induces Efficient mRNA Knockdown in Animal Embryos Developmental Cell. 2020.	Plasmid for bacterial expression and purification of RfxCas13d protein pET-28b-Cas9-His

Purify

A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA and its associated proteins, RNA, and DNA. The enCHIP system uses an anti-FLAG antibody to immunoprecipitate FLAG-tagged Cas9. Design your gRNA sequence to direct dCas9 to a specific locus, avoiding known transcription factor and other protein binding sites.

	Plasmid	Gene/Insert	Promoter	PI	Publication
	3xFLAG-dCas9/p-bacteria	3xFLAG-dCas9	pLtetO-1	Fujii	Efficient isolation of specific genomic regions and identification of associated proteins by engineered DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR. Biochem Biophys Res Commun. 2013 Aug 11. pii: S0006-291X(13)01329-6. doi: 10.1016/j.bbrc.2013.08.013.

Empty gRNA Expression Vectors

Select a gRNA expression plasmid based on factors such as selectable marker or cloning method. When using CRISPR, you will need to express both a Cas protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein act as all-in-one vectors, but their function is often limited to a single category (cut, nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with a wide variety of Cas-containing plasmids.

	gRNA Plasmid	Promoter	Cloning Enzyme(s)	Validated In	Resistance	Co-expressed Cas9	Depositing lab
Cas9 species = S. pyogenes (PAM = NGG)
	pCRISPR		BsaI	E. coli, S. pneumoniae	Kanamycin	none, need Cas9 plasmid	Marraffini
	pCas9		BsaI	E. coli, S. pneumoniae	Chloramphenicol	yes, cut	Marraffini
	pgRNA-bacteria	BBa_J23119	SpeI + HindIII		Ampicillin	none, need Cas9 plasmid	Qi