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CRISPR Plasmids: Plants

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CRISPR Resources

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

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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 Selectable Marker PI Publication Hidden Extra Search Info
pK7WGF2::hCas9hCas9 (Synthetic)Kanamycin Kamoun Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease. Nat Biotechnol. 2013 Aug 8;31(8):691-3. doi: 10.1038/nbt.2655. Expesses the human codon usage Cas9 nuclease (Mali et al. Science 339, 823-826, 2013) with an N-terminal GFP tag from the 35S promoter in the plant tissue pK7WGF2
pICH41308::hCas9hCas9 (Synthetic) Kamoun Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system. Plant Methods. 2013 Oct 11;9(1):39. Level 0 hCas9 module pICH41308
pICH47742::2x35S-5’UTR-hCas9(STOP)-NOST35Sp::hCas9 (Synthetic) Kamoun Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system. Plant Methods. 2013 Oct 11;9(1):39. Level 1 hCas9 module pICH47742
pAGM4723::AtU6p::sgRNA2-2x35S-5′UTR::Cas9::NOST-AtU6p::sgRNA1sgRNA_PDS2-Cas9-sgRNA_PDS1 (Synthetic) Kamoun Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas system. Plant Methods. 2013 Oct 11;9(1):39. Level 2 construct pAGM4723
pBUN4113×FLAG-NLS-zCas9-NLS (Synthetic), gRNA scaffold (Synthetic)Bar Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (OsU3 promoter), Bar resistance pGreen-like binary vector
pHSN4013×FLAG-NLS-zCas9-NLS (Synthetic), gRNA scaffold (Synthetic)Hygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Hyg resistance pGreen-like binary vector
pRGE31 Yang RNA-Guided Genome Editing in Plants Using a CRISPR-Cas System. Mol Plant. 2013 Nov;6(6):1975-83. doi: 10.1093/mp/sst119. Epub 2013 Aug 17. Expressing sgRNA and Cas9 in plants. The sgRNA was controlled by rice snoRNA U3 promoter pUGW11
pRGEB31Hygromycin Yang RNA-Guided Genome Editing in Plants Using a CRISPR-Cas System. Mol Plant. 2013 Nov;6(6):1975-83. doi: 10.1093/mp/sst119. Epub 2013 Aug 17. Binary vector derived from pRGE31. Deliver sgRNA and Cas9 into plant. by agrobacterium mediated transformation. pCAMBIA1300
HBT-pcoCas9pcoCas9 (Synthetic) Sheen Multiplex and homologous recombination-mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9. Nat Biotechnol. 2013 Aug;31(8):688-91. doi: 10.1038/nbt.2654. transient expression of pcoCas9 gene in plant cells HBT-FLAG
pFGC-pcoCas935SPPDK:pcoCas9:NOS cassette (Synthetic)Basta Sheen Sheen lab CRISPR plasmids (unpublished) A binary plasmid containing 35SPPDK:pcoCas9:NOS cassette for plant expression of Cas9 and multiple cloning sites for insertion of guide RNA pFGC-RCS
pJIT163-2NLSCas9Cas9 (Other) Gao Targeted genome modification of crop plants using a CRISPR-Cas system. Nat Biotechnol. 2013 Aug;31(8):686-8. doi: 10.1038/nbt.2650. Expression of rice codon-optimized Cas9 in plant cells pJIT163
p201N Cas9Cas9 (Synthetic), nptII (Other)G418, kanamycin Parrott Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 2015;15:16 Cas9 driven by double 35S, nptII for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle pPZP
p201H Cas9Cas9 (Synthetic), hph (Other)Hygromycin Parrott Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 2015;15:16 Cas9 driven by double 35S, hygromycin resistance for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle pPZP
p201B Cas9Cas9 (Synthetic), BAR (Other)Basta Parrott Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 2015;15:16 Cas9 driven by double 35S, BAR for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle pPZP
p201G Cas9Cas9 (Synthetic), sGFP (Synthetic)GFP Parrott Targeted genome modifications in soybean with CRISPR/Cas9. BMC Biotechnology. 2015;15:16 Cas9 driven by double 35S, GFP for plant selection, I-PpoI site to accept gRNA from pUC gRNA Shuttle pPZP
Cas9 MDC123Glycine Max codon optimized Cas9 (Other)Basta Stupar CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a new web-tool and a modified Cas9 enzyme. GM Crops Food. 2015;6(4):243-52. doi: 10.1080/21645698.2015.1106063. 2x35S promoting Glycing Max codon optimized Cas9 with bar plant selection pMDC123
Cas9 MDC32Glycine Max codon optimized Cas9 (Other)Hygromycin Stupar CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a new web-tool and a modified Cas9 enzyme. GM Crops Food. 2015;6(4):243-52. doi: 10.1080/21645698.2015.1106063. 2x35S promoting Glycing Max codon optimized Cas9 with hygromycin plant selection pMDC32
G10 Cas9 MDC123Glycine Max codon optimized Cas9 (Other)Basta Stupar CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a new web-tool and a modified Cas9 enzyme. GM Crops Food. 2015;6(4):243-52. doi: 10.1080/21645698.2015.1106063. G10 promoting Glycing Max codon optimized Cas9 (N and C terminus NLS) with bar plant selection pMDC123
pDe-CAS9Cas9 (Arabidopsis thaliana)Basta Puchta Both CRISPR/Cas-based nucleases and nickases can be used efficiently for genome engineering in Arabidopsis thaliana. Plant J. 2014 Jul;79(2):348-59. doi: 10.1111/tpj.12554. Epub 2014 Jun 17. Binary expression vector with A.th. codon-optimized Cas9 and Gateway destination sequence pPZP221
pCAS9-TPCCas9 (Arabidopsis thaliana)Basta Puchta Both CRISPR/Cas-based nucleases and nickases can be used efficiently for genome engineering in Arabidopsis thaliana. Plant J. 2014 Jul;79(2):348-59. doi: 10.1111/tpj.12554. Epub 2014 Jun 17. Binary expression vector with A.th. codon-optimized Cas9 for conventional cloning pPZP221
pBUE411gRNA scaffold (Synthetic), zCas9 (Other)Basta Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (OsU3 promoter), Bar resistance pCambia
pHSE401gRNA scaffold (Synthetic), zCas9 (Other)Hygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Hyg resistance pCambia
pKSE401gRNA scaffold (Synthetic), zCas9 (Other)Neomycin (select with G418) Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Kan resistance pCambia
pHUE411gRNA scaffold (Synthetic), zCas9 (Other)Hygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (OsU3 promoter), Hyg resistance pCambia
pBUN421gRNA scaffold (Synthetic), zCas9 (Other)Basta Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses 3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence (TaU3 promoter), Bar resistance pGreen-like
pRGEB32Hygromycin Yang Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice ubiquitin promoter for Agrobacterium-mediated transformation. pRGEB31
pRGE32 Yang Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice ubiquitin promoter for transient expression in protoplast pRGE31
pYLCRISPR/Cas9Pubi-HCas9 (Other)Hygromycin Liu A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. expression of Cas9 in plants, maize ubiquitin promoter (Pubi), hygro selection pCAMBIA1300
pYLCRISPR/Cas9Pubi-BCas9 (Other)Basta Liu A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. expression of Cas9 in plants, maize ubiquitin promoter (Pubi), basta selection pCAMBIA1300
pYLCRISPR/Cas9P35S-HCas9 (Other)Hygromycin Liu A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. expression of Cas9 in plants, cauliflower mosaic virus 35S promoter (P35S), hygro selection pCAMBIA1300
pYLCRISPR/Cas9P35S-BCas9 (Other)Basta Liu A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. expression of Cas9 in plants, cauliflower mosaic virus 35S promoter (P35S), basta selection pCAMBIA1300
pYLCRISPR/Cas9P35S-NCas9 (Other)Neomycin (select with G418) Liu A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. expression of Cas9 in plants, cauliflower mosaic virus 35S promoter (P35S), Neo selection pCAMBIA1300
pEGB2alpha2 35s:hCas9:tNos (GB0639)hCas9 (Other) Orzaez GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. Plant Physiol. 2013 Jul;162(3):1618-31. Transcriptional unit for (human codon optimized) Cas9 plant expression driven by the 35S promoter pDGB2alpha2
pICSL11056Promoter/5UTR: ZmUbi + CDS:SpCas9 + 3UTR/terminator:35s (Other) Patron Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease. Genome Biol. 2015 Nov 30;16(1):258. doi: 10.1186/s13059-015-0826-7. Level 1 Golden Gate Cassette: Cas9 expression cassette for monocotyledonous plants pICH47742 (AddGene #47742)
pICSL11060Promoter/5UTR: CsVMV+ CDS:SpCas9 + 3UTR/terminator:35s (Other) Patron Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease. Genome Biol. 2015 Nov 30;16(1):258. doi: 10.1186/s13059-015-0826-7. Level 1 Golden Gate Cassette: Cas9 expression cassette for plants (exemplified in Brassica oleracea) pICH47742 (AddGene #47742)
pYPQ150pcoCas9 (plant codon-optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with pcoCas9 (Plant codon optimized) pNJB91
pYPQ154AteCas9 (Arabidopsis codon optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with AteCas9 (Arabidopsis codon optimized) pYPQ185-linker2
pYPQ158hSpCas9 (Human codon optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with hSpCas9 (Human codon optimized) pNJB185-linker6
pYPQ167Cas9p (Plant codon-optimized; high GC content at 5 prime region) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with Cas9p pYPQ185-linker2
pTC217Nuclease (Cas9/sgRNA) + Donor + GVR (Other)kanamycin Voytas High-frequency, precise modification of the tomato genome. Genome Biol. 2015 Nov 6;16(1):232. doi: 10.1186/s13059-015-0796-9. Cas9/sgRNA targeting ANT1 locus, donor molecule with 5' homology arm-Pnos:NptII-35S:ANT13' homology arm as a Gemini Viral Replicon based on Bean Yellow Dwarf Virus; sgRNA= 1b pLSLR
pTC223Nuclease (Cas9/sgRNA) + Donor + GVR (Other)kanamycin Voytas High-frequency, precise modification of the tomato genome. Genome Biol. 2015 Nov 6;16(1):232. doi: 10.1186/s13059-015-0796-9. Cas9/sgRNA targeting ANT1 locus, donor molecule with 5' homology arm-Pnos:NptII-35S:ANT13' homology arm as a Gemini Viral Replicon based on Bean Yellow Dwarf Virus; sgRNA= 7 pLSLR
pHEE401sgRNA scaffold (Synthetic), zCas9 (Other)Hygromycin Chen Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. Egg cell-specific promoter-controlled expression of 3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target sequence (U6-26 promoter), Hyg resistance pCambia
pHEE401EgRNA scaffold (Synthetic), zCas9 (Other)Hygromycin Chen Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. Egg cell-specific promoter-controlled expression of 3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target sequence (U6-26 promoter), Hyg resistance pCambia
pHEE2E-TRIsgRNA targeting TRY and CPC genes (Synthetic), sgRNA targeting ETC2 gene (Synthetic), zCas9 (Other)Hygromycin Chen Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. Egg cell-specific promoter-controlled expression 3×FLAG-NLS-zCas9-NLS and two sgRNAs targeting three genes: ETC2, TRY, and CPC pCambia
pMpGE010Hygromycin Kohchi Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. PLoS One. 2018 Oct 31;13(10):e0205117. doi: 10.1371/journal.pone.0205117. eCollection 2018. For CRISPR/Cas9 genome editing in Marchantia polymorpha pMpGWB103
pMpGE011Chlorsulfuron Kohchi Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. PLoS One. 2018 Oct 31;13(10):e0205117. doi: 10.1371/journal.pone.0205117. eCollection 2018. For CRISPR/Cas9 genome editing in Marchantia polymorpha pMpGWB303
pEGB2Ω2_SF-35s:hCas9:tNos (GB1103)hCas9 (Other) Orzaez A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. eCollection 2016. Transcriptional unit for (human codon optimized) Cas9 plant expression driven by the 35S promoter. Specially conceived to be linked with the TU of the kanamycin resistance gene GB1181. pDGB2omega2
pBAtCBasta Kim A simple, flexible and high-throughput cloning system for plant genome editing via CRISPR-Cas system. J Integr Plant Biol. 2016 Mar 4. doi: 10.1111/jipb.12474. Plant expression of Cas9 and gRNA empty backbone; Basta resistance pB2GW7
  • Tag / Fusion Protein
    • NLS-HA (C terminal on insert)
    		•
    pHAtCHygromycin Kim A simple, flexible and high-throughput cloning system for plant genome editing via CRISPR-Cas system. J Integr Plant Biol. 2016 Mar 4. doi: 10.1111/jipb.12474. Plant expression of Cas9 and gRNA empty backbone; Hygro Resistance pH2GW7
    pHDE-35S-Cas9-mCherryHygromycin ; mCherry fluorescence Zhao An effective strategy for reliably isolating heritable and Cas9-free Arabidopsis mutants generated by CRISPR/Cas9-mediated genome editing. Plant Physiol. 2016 May 15. pii: pp.00663.2016. For CRISPR/Cas9 mediated gene editing in Arabidopsis; provides a visual screen for Cas9-free plants pHDE
    pHDE-35S-Cas9-mCherry-UBQHygromycin ; mCherry fluorescence Zhao An effective strategy for reliably isolating heritable and Cas9-free Arabidopsis mutants generated by CRISPR/Cas9-mediated genome editing. Plant Physiol. 2016 May 15. pii: pp.00663.2016. For CRISPR/Cas9 mediated gene editing in Arabidopsis; provides a visual screen for Cas9-free plants. Enable production of two gRNAs pHDE
    pKIR1.1human-codon-optimized SpCas9 (Other)Hygromycin ; TagRFP in seeds Higashiyama pKAMA-ITACHI vectors for highly efficient CRISPR/Cas9-mediated gene knockout in Arabidopsis thaliana. Plant Cell Physiol. 2016 Nov 17. pii: pcw191. CRISPR/Cas9 in Arabidopsis with seed a fluorescent reporter pFAST-R01
    pKI1.1human-codon-optimized SpCas9 (Other) Higashiyama pKAMA-ITACHI vectors for highly efficient CRISPR/Cas9-mediated gene knockout in Arabidopsis thaliana. Plant Cell Physiol. 2016 Nov 17. pii: pcw191. CRISPR/Cas9 for Arabidopsis in pENTR/D-TOPO pENTR/D-TOPO
    pKI1.1Rhuman-codon-optimized SpCas9 (Other)Hygromycin ; TagRFP in seeds Higashiyama pKAMA-ITACHI vectors for highly efficient CRISPR/Cas9-mediated gene knockout in Arabidopsis thaliana. Plant Cell Physiol. 2016 Nov 17. pii: pcw191. CRISPR/Cas9 in Arabidopsis with seed a fluorescent reporter. pFAST-R01
    pAGM4723:TpCC_UreaseCas9 (Other), Urease-targeting gRNA 1 (Other), Urease-targeting gRNA 2 (Other)Nourseothricin (Nat) Mock Editing of the urease gene by CRISPR-Cas in the diatom Thalassiosira pseudonana Plant Methods 2016, 12:49 Golden Gate Level 2 construct encoding Cas9-YFP and 2 urease-targeting gRNAs pAGM4723
    pICH47742:FCP:Cas9YFPCas9 (Other) Mock Editing of the urease gene by CRISPR-Cas in the diatom Thalassiosira pseudonana Plant Methods 2016, 12:49 Golden Gate Level 1 cassette encoding a Cas9-YFP fusion under the FCP promoter pICH47742
    pYPQ220 (AsCpf1)AsCpf1 (Other) Qi A CRISPR-Cpf1 system for efficient genome editing and transcriptional repression in plants. Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18. AsCpf1 Gateway entry plasmid pYPQ167
    pYPQ230 (LbCpf1)LbCpf1 (Other) Qi A CRISPR-Cpf1 system for efficient genome editing and transcriptional repression in plants. Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18. LbCpf1 Gateway entry plasmid pYPQ167
    pKIR1.0human-codon-optimized SpCas9 (Other)TagRFP in seeds Higashiyama pKAMA-ITACHI vectors for highly efficient CRISPR/Cas9-mediated gene knockout in Arabidopsis thaliana. Plant Cell Physiol. 2016 Nov 17. pii: pcw191. CRISPR/Cas9 in Arabidopsis with seed a fluorescent reporter. pFAST-R01
    pUB-Cas9Cas9 (Synthetic)Hygromycin Weber An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis thaliana. Front Plant Sci. 2017 Jan 24;8:39. doi: 10.3389/fpls.2017.00039. eCollection 2017. For ubiquitous expression of Cas9 in planta. sgRNA can be introduced as described in Hahn et al. (2017) pKB65
    [email protected]sgRNA against GLABROUS1 (Synthetic)Hygromycin Weber An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis thaliana. Front Plant Sci. 2017 Jan 24;8:39. doi: 10.3389/fpls.2017.00039. eCollection 2017. Disruption of GLABROUS1 gene in Arabidopsis using CRISPR/Cas9 pUB-Cas9
    pTX168Hygromycin Voytas A Single Transcript CRISPR-Cas9 System for Efficient Genome Editing in Plants. Mol Plant. 2016 Jul 6;9(7):1088-91. doi: 10.1016/j.molp.2016.05.001. Epub 2016 May 19. Express STU (Single Transcriptional Unit) Cas9 with CaMV 35s promoter in plants pCAMBIA1300
  • Tag / Fusion Protein
    • SV40 NLS (C terminal on insert)
    		•
    pTX171Hygromycin Voytas A Single Transcript CRISPR-Cas9 System for Efficient Genome Editing in Plants. Mol Plant. 2016 Jul 6;9(7):1088-91. doi: 10.1016/j.molp.2016.05.001. Epub 2016 May 19. Express STU (Single Transcriptional Unit) Cas9 with CaMV 35s promoter in plants pTX168
  • Tag / Fusion Protein
    • SV40 NLS (C terminal on insert)
    		•
    pTX172Hygromycin Voytas A Single Transcript CRISPR-Cas9 System for Efficient Genome Editing in Plants. Mol Plant. 2016 Jul 6;9(7):1088-91. doi: 10.1016/j.molp.2016.05.001. Epub 2016 May 19. Express STU (Single Transcriptional Unit) Cas9 with Maize Ubiquitin1 promoter in plants pTX171
  • Tag / Fusion Protein
    • SV40 NLS (C terminal on insert)
    		•
    pTX176Hygromycin Voytas A Single Transcript CRISPR-Cas9 System for Efficient Genome Editing in Plants. Mol Plant. 2016 Jul 6;9(7):1088-91. doi: 10.1016/j.molp.2016.05.001. Epub 2016 May 19. Express estrodial-inducible STU (Single Transcriptional Unit) Cas9 in plants pTX171
  • Tag / Fusion Protein
    • SV40 NLS (C terminal on insert)
    		•
    pJG80TaCas9 (Other) Voytas High-efficiency gene targeting in hexaploid wheat using DNA replicons and CRISPR/Cas9. Plant J. 2016 Dec 10. doi: 10.1111/tpj.13446. Entry vector containing TaCas9-nos terminator between attL1 and attR5 sites unknown
    pMOD_A0108 AtCas9 (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: EC1.2, Gene: AtCas9, Terminator: HSP pMOD_A1001
    pMOD_A0502 Csy4-P2A-AtCas9 (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: AtUbi10, Gene: Csy4-P2A-AtCas9, Terminator: HSP pMOD_A1001
    pMOD_A0508 Csy4-P2A-AtCas9 (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: EC1.2, Gene: Csy4-P2A-AtCas9, Terminator: HSP pMOD_A1001
    pMOD_A0901 TREX2-P2A-AtCas9 (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: 35S, Gene: TREX2-P2A-AtCas9, Terminator: HSP pMOD_A1001
    pMOD_A0908 TREX2-P2A-AtCas9 (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: EC1.2, Gene: TREX2-P2A-AtCas9, Terminator: HSP pMOD_A1001
    pMOD_A1110 TaCas9 (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: ZmUbi, Gene: TaCas9, Terminator: HSP pMOD_A1001
    pMOD_A1111 TaCas9 (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: OsAct1, Gene: TaCas9, Terminator: HSP pMOD_A1001
    pMOD_A1510 Csy4-P2A-TaCas9 (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: ZmUbi, Gene: Csy4-P2A-TaCas9, Terminator: HSP pMOD_A1001
    pMOD_A1511 Csy4-P2A-TaCas9 (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: OsAct1, Gene: Csy4-P2A-TaCas9, Terminator: HSP pMOD_A1001
    pDIRECT_21A Engineering Reagent: 35S:AtCas9 + AtU6:gRNA (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: 35S:AtCas9 + AtU6:gRNA, Plant Selection: 2x35S:hpt II pTRANS_210
    pDIRECT_21C Engineering Reagent: 35S:Csy4-P2A-AtCas9 + CmYLCV:gRNAs with Csy4 spacers (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: 35S:Csy4-P2A-AtCas9 + CmYLCV:gRNAs with Csy4 spacers , Plant Selection: 2x35S:hpt II pTRANS_210d
    pDIRECT_22A Engineering Reagent: 35S:AtCas9 + AtU6:gRNA (Synthetic)Kanamycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: 35S:AtCas9 + AtU6:gRNA, Plant Selection: 2x35S:npt II pTRANS_220
    pDIRECT_22C Engineering Reagent: 35S:Csy4-P2A-AtCas9 + CmYLCV:gRNAs with Csy4 spacers (Synthetic)Kanamycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: 35S:Csy4-P2A-AtCas9 + CmYLCV:gRNAs with Csy4 spacers , Plant Selection: 2x35S:npt II pTRANS_220d
    pDIRECT_23A Engineering Reagent: 35S:AtCas9 + AtU6:gRNA (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: 35S:AtCas9 + AtU6:gRNA, Plant Selection: 2x35S:bar pTRANS_230
    pDIRECT_23C Engineering Reagent: 35S:Csy4-P2A-AtCas9 + CmYLCV:gRNAs with Csy4 spacers (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: 35S:Csy4-P2A-AtCas9 + CmYLCV:gRNAs with Csy4 spacers , Plant Selection: 2x35S:bar pTRANS_230d
    pDIRECT_25F Engineering Reagent: ZmUbi:TaCas9 + TaU6:gRNA (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: ZmUbi:TaCas9 + TaU6:gRNA, Plant Selection: PvUbi2:hpt II pTRANS_250d
    pDIRECT_25H Engineering Reagent: ZmUbi:Csy4-P2A-TaCas9 + PvUbi1:gRNAs with Csy4 spacers (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: ZmUbi:Csy4-P2A-TaCas9 + PvUbi1:gRNAs with Csy4 spacers, Plant Selection: PvUbi2:hpt II pTRANS_250d
    pDIRECT_26H Engineering Reagent: ZmUbi:Csy4-P2A-TaCas9 + PvUbi1:gRNAs with Csy4 spacers (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: ZmUbi:Csy4-P2A-TaCas9 + PvUbi1:gRNAs with Csy4 spacers, Plant Selection: PvUbi2:bar pTRANS_260d
    pAtCas9_1 AtCas9 (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9, Plant Selection: 2x35S:hpt II pCAMBIA
    pAtCas9_3 AtCas9 (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9, Plant Selection: 2x35S:bar pCAMBIA
    pTaCas9_5 TaCas9 (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: TaCas9, Plant Selection: PvUbi2:hpt II pCAMBIA
    pTaCas9_6 TaCas9 (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: TaCas9, Plant Selection: PvUbi2:bar pCAMBIA
    pDIRECT_10A35S:AtCas9 + AtU6:gRNA (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: non-T-DNA, Engineering Reagent: 35S:AtCas9 + AtU6:gRNA, Plant Selection: none pTRANS_100
    pKEE401Neomycin (select with G418) Chen Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. CRISPR/Cas9-mediated genome editing in Arabidopsis. Contains Cas9 and empty gRNA scaffold, Kan resistance pCambia
    pXEE401B Chen Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. CRISPR/Cas9-mediated genome editing in Arabidopsis. Contains Cas9 and empty gRNA scaffold, Bar resistance pCambia
    pXEE401BG Chen Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0. CRISPR/Cas9-mediated genome editing in Arabidopsis, Bar and glyphosate-resistance pCambia
    pSC10 Stupar CRISPR/Cas9 and TALENs generate heritable mutations for genes involved in small RNA processing of Glycine max and Medicago truncatula. Plant Biotechnol J. 2017 Oct 31. doi: 10.1111/pbi.12857. pAH595-gRNA 2xplex entry cassette pENTR top
    pSC12 Stupar CRISPR/Cas9 and TALENs generate heritable mutations for genes involved in small RNA processing of Glycine max and Medicago truncatula. Plant Biotechnol J. 2017 Oct 31. doi: 10.1111/pbi.12857. pNB184-Cas9 entry cassette pENTR top
    pYLCRISPR/Cas9pUbi-NCas9 (Synthetic) Liu A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. Expression of Cas9 in plants, ZmUbi promoter, Kanamycin selection pCAMBIA1300
    pJKW0458 Weng pJKW0457 (unpublished) p35S:CAS9 pEarleyGate100
    pJKW0474 Weng pJKW0457 (unpublished) pCLV3:Cas9 pEarleyGate100
    pMpGE006Atco-Cas9-Pea3ter (Synthetic)Hygromycin Sugano Efficient CRISPR/Cas9-based genome editing and its application to conditional genetic analysis in Marchantia polymorpha. PLoS One. 2018 Oct 31;13(10):e0205117. doi: 10.1371/journal.pone.0205117. eCollection 2018. Cas9 expression in M.polymorpha pMpGWB103
    pYPQ239FnCpf1 (Other) Qi Plant Genome Editing Using FnCpf1 and LbCpf1 Nucleases at Redefined and Altered PAM Sites. Mol Plant. 2018 Jul 2;11(7):999-1002. doi: 10.1016/j.molp.2018.03.008. Epub 2018 Mar 20. FnCpf1 Gateway entry plasmid pYPQ167
  • Tags / Fusion Proteins
    • NLS (N terminal on insert)
    • NLS (C terminal on insert)
    		•
    pORFE0001AtCAS9 (Other) Chevalier Chevalier Lab plasmids (unpublished) AtCas9 module CDS1 pICH41308
    pORFE1001AtCAS9 (Other) Chevalier Chevalier Lab plasmids (unpublished) Overexpression of the AtCAS9 in plant under 2x35S promoter pICH47742
    pMH-Cas9-gateCas9 (Other)Hygromycin Bezanilla Efficient and modular CRISPR-Cas9 vector system for Physcomitrella patens. Plant Direct. 2019 Sep 12;3(9):e00168. doi: 10.1002/pld3.168. eCollection 2019 Sep. Gateway destination vector encoding Maize ubiquitin promoter driving Cas9 and 35S promoter driving hygromycin for selection in plants. pGEM-T easy
    pMK-Cas9-gateCas9 (Other)Neomycin (select with G418) Bezanilla Efficient and modular CRISPR-Cas9 vector system for Physcomitrella patens. Plant Direct. 2019 Sep 12;3(9):e00168. doi: 10.1002/pld3.168. eCollection 2019 Sep. Gateway destination vector encoding Maize ubiquitin promoter driving Cas9 and 35S promoter driving aminoglycoside phosphotransferase for G418 selection in plants. pGEM-T easy
    pZeo-Cas9-gateCas9 (Other)Zeocin Bezanilla Efficient and modular CRISPR-Cas9 vector system for Physcomitrella patens. Plant Direct. 2019 Sep 12;3(9):e00168. doi: 10.1002/pld3.168. eCollection 2019 Sep. Gateway destination vector encoding Maize ubiquitin promoter driving Cas9 and 35S promoter driving Zeocin for selection in plants. pGEM-T easy
    pICSL90016BsaI:AATG:Cas9_2:GCTT:BsaI (Synthetic) Jones Optimization of T-DNA architecture for Cas9-mediated mutagenesis in Arabidopsis bioRxiv 419952 Cas9_2 Golden Gate Level 0 pICH41308
    BCJJ180BsaI:AATG:Cas9_4:GCTT:BsaI (Synthetic) Jones Optimization of T-DNA architecture for Cas9-mediated mutagenesis in Arabidopsis bioRxiv 419952 Cas9_4 Golden Gate Level 0 pICH41308
    pICSL90005 SpCas9-h (no stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, SpCas9-h (no stop codon) pAGM1287
    pEPOR0SP0013SpCas9-p (with stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, SpCas9-p pICH41308
    pEPOR0SP0009SpCas9-p (no stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, SpCas9-p (no stop codon) pAGM1287
    pEPOR0SP0001SpCas9-DE (no stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, SpCas9-DE (no stop codon) pAGM1287
    pEPOR0SP0014SpCas9-KA (with stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, SpCas9-KA pICH41308
    pEPOR0SP0015SpCas9-KA (no stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, SpCas9-KA (no stop codon) pAGM1287
    pEPOR0SP0016Sp-eCas9 1.0 (with stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, Sp-eCas9 1.0 pICH41308
    pEPOR0SP0017Sp-eCas9 1.0 (no stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, Sp-eCas9 1.0 (no stop codon) pAGM1287
    pEPOR0SP0018Sp-eCas9 1.1 (with stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, Sp-eCas9 1.1 pICH41308
    pEPOR0SP0019Sp-eCas9 1.1 (no stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, Sp-eCas9 1.1 (no stop codon) pAGM1287
    pEPOR0SP0011Sp-xCas9 3.7 (no stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, Sp-xCas9 3.7 (no stop codon) pAGM1287
    pEPOR0SP0020SaCas9 (with stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, SaCas9 pICH41308
    pEPOR0SP0021SaCas9 (no stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, SaCas9 (no stop codon) pAGM1287
    pEPOR0SP0012eSaCas9 (no stop codon) (Other) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level0 Golden Gate part: CDS, eSaCas9 (no stop codon) pAGM1287
    pICSL110232x35S+5'UTR OMEGA (pICH51288) + SpCas9-h (no stop codon)(pICSL90005) + YFP (pICSL50005)+ Nos (pICH41421) (Synthetic) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level 1 Golden Gate Cassette: Cas9 expression cassette for dicotyledonous plants pICH47742
    pEPOR1CB00022x35S+5'UTR OMEGA (pICH51288) + SpCas9-p (no stop codon)(pEPOR0SP0009) + YFP (pICSL50005)+ Nos (pICH41421) (Synthetic) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level 1 Golden Gate Cassette: Cas9 expression cassette for dicotyledonous plants pICH47742
    pEPOR1CB00092x35S+5'UTR OMEGA (pICH51288) + SpCas9-KA (no stop codon)(pEPOR0SP0015) + YFP (pICSL50005)+ Nos (pICH41421) (Synthetic) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level 1 Golden Gate Cassette: Cas9-K855A expression cassette for dicotyledonous plants pICH47742
    pEPOR1CB00112x35S+5'UTR OMEGA (pICH51288) + Sp-eCas9 1.1 (no stop codon) (pEPOR0SP0019 + YFP (pICSL50005) + Nos (pICH41421) (Synthetic) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level 1 Golden Gate Cassette: eCas9 1.1 expression cassette for dicotyledonous plants pICH47742
    pEPOR1CB01122x35S+5'UTR OMEGA (pICH51288) + Sp-xCas9 3.7 (no stop codon) (pEPOR0SP0011) + YFP (pICSL50005) + Nos (pICH41421) (Synthetic) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level 1 Golden Gate Cassette: xCas9 3.7 expression cassette for dicotyledonous plants pICH47742
    pEPOR1CB00152x35S+5'UTR OMEGA (pICH51288) + SaCas9 (no stop codon) (pEPOR0SP0012) + YFP (pICSL50005) + Nos (pICH41421) (Synthetic) Patron Comparison of efficiency and specificity of CRISPR-associated (Cas) nucleases in plants: An expanded toolkit for precision genome engineering. bioRxiv 422766 Level 1 Golden Gate Cassette: SaCas9 expression cassette for dicotyledonous plants pICH47742
    p221z-CAS9p-t35sCAS9p (Other) Mähönen Plants CRISPR plasmids (unpublished) Entry clone containing CAS9p and 35S terminator. Used to make CRISPR construct . For use in plants and compatible with the MultiSite Gateway system pDONR-221z
    p221z-CAS9p-TagRFP-t35sCAS9p-TagRFP (Other) Mähönen Plants CRISPR plasmids (unpublished) Entry clone containing CAS9p-TagRFP and 35S terminator. Used to make CRISPR construct . For use in plants and compatible with the MultiSite Gateway system pDONR-221z
    pYPQ150-x3.7pcoCas9-x3.7 Qi Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-NG. Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 10.1016/j.molp.2019.03.011. Gateway entry vector with pcoCas9-x3.7 (Plant codon optimized) pNJB91
    pGEL062Hygromycin Zhang Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-NG. Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 10.1016/j.molp.2019.03.011. For genome editing in plant by SpCas9 wild type pZHY988
    pGEL063Hygromycin Zhang Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-NG. Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 10.1016/j.molp.2019.03.011. For genome editing in plant by SpCas9 variant(SpCas9-NG) pZHY988
    pGEL065Hygromycin Zhang Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-NG. Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 10.1016/j.molp.2019.03.011. For genome editing in plant by SpCas9 variant(SpCas9-NGv1) pZHY988
    pFH13SpCas9, wheat codon optimized Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 SpCas9, wheat codon optimized pICH41308
    pFH14SaCas9, Arabidopsis codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 SaCas9, Arabidopsis codon optimized pICH41308
    pFH15SaCas9, Arabidopsis codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 StCas9, Arabidopsis codon optimized pICH41308
    pFH16FnCas12a, rice codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 FnCas12a, rice codon optimized pICH41308
    pFH17LbCas12a, rice codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 LbCas12a, rice codon optimized pICH41308
    pFH22xCas9 3.7 (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 xCas9 3.7 pICH41308
    pFH32SpCas9-NG, human codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 SpCas9-NG, human codon optimized pICH41308
    pFH46FnCas12a, human codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 FnCas12a, human codon optimized pICH41308
    pFH47LbCas12a, human codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 LbCas12a, human codon optimized pICH41308
    pFH48Csy4 (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 Csy4 pICH41308
    pFH76ScCas9, wheat codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 ScCas9, wheat codon optimized pICH41308
    pRGEB32-BARBasta ; BAR Hunter CRISPR/Cas9 vector optimized for Agrobacterium-mediated transformation of maize (with BAR selectable marker) (modified from pRGEB32) (unpublished) For CRISPR/Cas9 delivery in maize. Expresses Cas9 with rice ubiquitin promoter, BAR with 35S promoter, and sgRNA/PTG with rice snoRNA U3 promoter (pol III). Can produce 1 or more gRNAs. pRGEB32
    pENTR_Wheat_live_Cas9Wheat_live_Cas9 (Synthetic) Kamoun Genome editing in wheat (unpublished) Gateway pENTR-plasmid to generate a binary construct for genome editing of the wheat homolog of a susceptibility gene of O. sativa. Encodes wheat optimized Cas9 and BpiI sites for sgRNA modules. pENTR4
    pICSL11085_GG_Wheat_live_Cas9_forWheat_live_Cas9 (Synthetic) Kamoun Genome editing in wheat (unpublished) Encodes a wheat codon optimized Cas9 module in forward direction for golden gate cloning of genome editing binary plasmids. pICH47742
    pICSL11086_GG_Wheat_live_Cas9_revWheat_live_Cas9 (Synthetic) Kamoun Genome editing in wheat (unpublished) Encodes a wheat codon optimized Cas9 module in forward direction for golden gate cloning of genome editing binary plasmids. pICH47742
    pFH23ZmUBIp::SpCas9 (wheat codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::SpCas9 (wheat codon optimized):NOSt pICH47742
    pFH522x35Sp::SpCas9 (Arabidopsis codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 2x35Sp::SpCas9 (Arabidopsis codon optimized):NOSt pICH47742
    pFH532x35Sp::SpCas9 (Arabidopsis codon optimized):pea3At (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 2x35Sp::SpCas9 (Arabidopsis codon optimized):pea3At pICH47742
    pFH54AtUBI10p::SpCas9 (Arabidopsis codon optimized):pea3At (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 AtUBI10p::SpCas9 (Arabidopsis codon optimized):pea3At pICH47742
    pFH58ZmUBIp::SaCas9 (Arabidopsis codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::SaCas9 (Arabidopsis codon optimized):NOSt pICH47742
    pFH59ZmUBIp::StCas9 (Arabidopsis codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::StCas9 (Arabidopsis codon optimized):NOSt pICH47742
    pFH60ZmUBIp::FnCas12a (rice codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::FnCas12a (rice codon optimized):NOSt pICH47742
    pFH61ZmUBIp::LbCas12a (rice codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::LbCas12a (rice codon optimized):NOSt pICH47742
    pFH68ZmUBIp::SpCas9-NG (human codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::SpCas9-NG (human codon optimized):NOSt pICH47742
    pFH80ZmUBIp::xCas9 3.7:NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::xCas9 3.7:NOSt pICH47742
    pFH81ZmUBIp::ScCas9 (wheat codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::ScCas9 (wheat codon optimized):NOSt pICH47742
    pARS3_MUbCAS9_GWHygromycin Schaller Type-B response regulators of rice play key roles in growth, development, and cytokinin signaling. Development. 2019 Jun 3. pii: dev.174870. doi: 10.1242/dev.174870. T-DNA binary vector for CRISPR/Cas9 gene editing in rice; has a Gateway cassette for cloning gRNA pARS3
  • Tag / Fusion Protein
    • HA
    • pARS3_MUbCAS9_MCHygromycin Nimchuk Type-B response regulators of rice play key roles in growth, development, and cytokinin signaling. Development. 2019 Jun 3. pii: dev.174870. doi: 10.1242/dev.174870. T-DNA binary vector for CRISPR/Cas9 gene editing in rice; has a MCS for cloning gRNA pARS3
  • Tag / Fusion Protein
    • HA
    • pFH24SpCas9, wheat codon optimized, version 2 (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 SpCas9, wheat codon optimized (version 2) pICH41308
    pFH25SpCas9, wheat codon optimized, version 3 (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 SpCas9, wheat codon optimized (version 3) pICH41308
    pFH66ZmUBIp::SpCas9 (wheat codon optimized, version 2):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::SpCas9 (wheat codon optimized, version 2):NOSt pICH47742
    pFH67ZmUBIp::SpCas9 (wheat codon optimized, version 3):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::SpCas9 (wheat codon optimized, version 3):NOSt pICH47742
    pG3H-U3Ub CRISPR/Cas9 (Synthetic) Chen A Novel Ternary Vector System United with Morphogenic Genes Enhances CRISPR/Cas Delivery in Maize. Plant Physiol. 2019 Sep 26. pii: pp.19.00767. doi: 10.1104/pp.19.00767. pGreen3 CRISPR/Cas9 pGreen3

    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 Gene/Insert Promoter Selectable Marker PI Publication Hidden Extra Search Info
    pMonAID_dCas9-PmCDA_Hyg_ALSSpCas9 (Other)2x35SHygromycin Kondo Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion. Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3833. Monocot Target-AID vector expressing rice-optimized dCas9-PmCDA1 with sgRNA targeting OsAls pZDgRNA_Cas9ver.2 (D10A)_HPT
    pMonAID_nCas9-PmCDA_Hyg_ALSSpCas9 (Other)2x35SHygromycin Kondo Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion. Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3833. Monocot Target-AID vector expressing rice-optimized nCas9-PmCDA1 with sgRNA targeting OsAls pZDgRNA_Cas9ver.2 (D10A)_HPT
    pDicAID_nCas9-PmCDA_NptII_DellaSpCas9 (Other)PcUbiKanamycin Kondo Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion. Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3833. Dicot Target-AID vector expressing dicot-optimized nCas9-PmCDA1 with sgRNA targeting SlDella pZD_OsU3gYSA_HolgerCas9_NPTII
    pDicAID_nCas9-PmCDA-2A-NptII_ETRSpCas9 (Other)PcUbiKanamycin Kondo Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion. Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3833. Dicot Target-AID vector expressing dicot-optimized nCas9-PmCDA1-2A-NptII with sgRNA targeting SlEtr pZD_OsU3gYSA_HolgerCas9_NPTII
    pHEE901Hygromycin Chen CRISPR/Cas9-mediated base-editing system efficiently generates gain-of-function mutations in Arabidopsis. Sci China Life Sci. 2017 Mar 16. doi: 10.1007/s11427-017-9021-5. CRISPR/Cas9-mediated base editing in Arabidopsis. Contains Cas9(D10A) and empty gRNA scaffold, HygR pCambia
    pHSE901Hygromycin Chen CRISPR/Cas9-mediated base-editing system efficiently generates gain-of-function mutations in Arabidopsis. Sci China Life Sci. 2017 Mar 16. doi: 10.1007/s11427-017-9021-5. CRISPR/Cas9-mediated base editing in dicots. Contains Cas9(D10A) under CaMV 35S promoter and empty gRNA scaffold, HygR pCambia
    pBSE901Neomycin (select with G418) Chen CRISPR/Cas9-mediated base-editing system efficiently generates gain-of-function mutations in Arabidopsis. Sci China Life Sci. 2017 Mar 16. doi: 10.1007/s11427-017-9021-5. CRISPR/Cas9-mediated base editing in dicots. Contains Cas9(D10A) under CaMV 35S promoter and empty gRNA scaffold, Basta resistance pCambia
    pXSE901BBasta Chen CRISPR/Cas9-mediated base-editing system efficiently generates gain-of-function mutations in Arabidopsis. Sci China Life Sci. 2017 Mar 16. doi: 10.1007/s11427-017-9021-5. CRISPR/Cas9-mediated base editing in dicots. Contains Cas9(D10A) under CaMV 35S promoter and empty gRNA scaffold, Bar resistance pCambia
    pXSE901BGBasta ; Glyphosate Chen CRISPR/Cas9-mediated base-editing system efficiently generates gain-of-function mutations in Arabidopsis. Sci China Life Sci. 2017 Mar 16. doi: 10.1007/s11427-017-9021-5. CRISPR/Cas9-mediated base editing in dicots. Contains Cas9(D10A) under CaMV 35S promoter and empty gRNA scaffold, Basta and glyphosate-resistance pCambia
    pB-dCas9-PBENLS-APOBEC1-XTEN-dCas9-UGI-NLSUbiBasta Gao Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat Biotechnol. 2017 May;35(5):438-440. doi: 10.1038/nbt.3811. Epub 2017 Feb 27. Expresses CRISPR base editor in maize pBUE411
    pB-nCas9-PBENLS-APOBEC1-XTEN-nCas9-UGI-NLSUbiBasta Gao Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat Biotechnol. 2017 May;35(5):438-440. doi: 10.1038/nbt.3811. Epub 2017 Feb 27. Expresses CRISPR base editor in maize pBUE411
    pdCas9-PBENLS-APOBEC1-XTEN-dCas9-UGI-NLSUbi Gao Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat Biotechnol. 2017 May;35(5):438-440. doi: 10.1038/nbt.3811. Epub 2017 Feb 27. Expresses CRISPR base editor in protoplasts pJIT163-Ubi-GFP
    pH-dCas9-PBENLS-APOBEC1-XTEN-dCas9-UGI-NLSUbiHygromycin Gao Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat Biotechnol. 2017 May;35(5):438-440. doi: 10.1038/nbt.3811. Epub 2017 Feb 27. Expresses CRISPR base editor in rice pHUE411
    pH-nCas9-PBENLS-APOBEC1-XTEN-nCas9-UGI-NLSUbiHygromycin Gao Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat Biotechnol. 2017 May;35(5):438-440. doi: 10.1038/nbt.3811. Epub 2017 Feb 27. Expresses CRISPR base editor in rice pHUE411
    pnCas9-PBENLS-APOBEC1-XTEN-nCas9-UGI-NLSUbi Gao Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat Biotechnol. 2017 May;35(5):438-440. doi: 10.1038/nbt.3811. Epub 2017 Feb 27. Expresses CRISPR base editor in protoplasts pJIT163-Ubi-GFP
    pJY-RpABERPS5A promoter (Arabidopsis thaliana), ABE7.10 (Other), U6-sgRNA cassetteHygromycin Kim Precision genome engineering through adenine base editing in plants. Nat Plants. 2018 Jun 4. pii: 10.1038/s41477-018-0178-x. doi: 10.1038/s41477-018-0178-x. binary vector for ABE expression in Arabidopsis pKI1.1R
    pJY-RpABE-FT_INFRPS5A promoter (Arabidopsis thaliana), ABE7.10 (Other), U6-sgRNA targeting FTHygromycin Kim Precision genome engineering through adenine base editing in plants. Nat Plants. 2018 Jun 4. pii: 10.1038/s41477-018-0178-x. doi: 10.1038/s41477-018-0178-x. binary vector for INF (Induced Neo-Functionalization) of FT in Arabidopsis pJY-RpABE
    pJY-RpABE-PDS3_IMSRPS5A promoter (Arabidopsis thaliana), ABE7.10 (Other), U6-sgRNA targeting PDS3Hygromycin Kim Precision genome engineering through adenine base editing in plants. Nat Plants. 2018 Jun 4. pii: 10.1038/s41477-018-0178-x. doi: 10.1038/s41477-018-0178-x. binary vector for IMS (Induced Mis-Splicing) of PDS3 in Arabidopsis pJY-RpABE
    pH-PABE-7-esgRNAwtTadA-TadA7.10-nCas9-3*NLS (Other) Gao Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. Targeted A to G in rice pCAMBIA1300
    pH-PABE-7-sgRNAwtTadA-TadA7.10-nCas9-3*NLS (Other) Gao Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. Targeted A to G in rice pCAMBIA1300
    PABE-1NLS-wtTadA-TadA7.10-nCas9-NLS (Other) Gao Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. Targeted A to G in wheat plants or monocotyledons protoplasts pJIT163
    PABE-2wtTadA-TadA7.10-nCas9-NLS (Other) Gao Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. Targeted A to G in wheat plants or monocotyledons protoplasts pJIT163
    PABE-3wtTadA-TadA7.10-NLS-nCas9-NLS (Other) Gao Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. Targeted A to G in wheat plants or monocotyledons protoplasts pJIT163
    PABE-4NLS-nCas9-wtTadA-TadA7.10-NLS (Other) Gao Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. Targeted A to G in wheat plants or monocotyledons protoplasts pJIT163
    PABE-5NLS-nCas9-NLS-wtTadA-TadA7.10-NLS (Other) Gao Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. Targeted A to G in wheat plants or monocotyledons protoplasts pJIT163
    PABE-6wtTadA-TadA7.10-nCas9-2*NLS (Other) Gao Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. Targeted A to G in wheat plants or monocotyledons protoplasts pJIT163
    PABE-7wtTadA-TadA7.10-nCas9-3*NLS (Other) Gao Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z. Targeted A to G in wheat plants or monocotyledons protoplasts pJIT163
    pYPQ255rAPOBEC1-pcoCas9D10A-UGI Qi Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-NG. Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 10.1016/j.molp.2019.03.011. Gateway entry vector with rAPOBEC1 and pcoCas9D10A for C-T base editing pNJB91
    pYPQ255-x3.7rAPOBEC1-pcoCas9D10A_x3.7-UGI Qi Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-NG. Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 10.1016/j.molp.2019.03.011. Gateway entry vector with rAPOBEC1 and pcoCas9D10A-x3.7 for C-T base editing pNJB91
    pYPQ256pcoCas9D10A-PmCDA1-UGI Qi Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-NG. Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 10.1016/j.molp.2019.03.011. Gateway entry vector with PmCDA1 and pcoCas9D10A for C-T base editing pNJB91
    pYPQ256-x3.7pcoCas9D10A_x3.7-PmCDA1-UGI Qi Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-NG. Mol Plant. 2019 Mar 27. pii: S1674-2052(19)30124-8. doi: 10.1016/j.molp.2019.03.011. Gateway entry vector with PmCDA1 and pcoCas9D10A-x3.7 for C-T base editing pNJB91
    pFH45nSpCas9-ABE7.10 base changer, human codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 nSpCas9-ABE7.10 base changer, human codon optimized pICH41308
    pFH55nSpCas9-PmCDA, Arabidopsis codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 nSpCas9-PmCDA, Arabidopsis codon optimized pICH41308
    pFH79A3A-PBE, wheat codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 A3A-PBE, wheat codon optimized pICH41308
    pFH92nScCas9-ABE7.10, wheat/human codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 nScCas9-ABE7.10, wheat/human codon optimized pICH41308
    pFH70ZmUBIp::nSpCas9-PmCDA (Arabidopsis codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::nSpCas9-PmCDA (Arabidopsis codon optimized):NOSt pICH47742
    pFH84ZmUBIp::A3A-PBE (wheat codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::A3A-PBE (wheat codon optimized):NOSt pICH47742
    pFH93ZmUBIp::nScCas9-ABE7.10 (wheat/human codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::nScCas9-ABE7.10 (wheat/human codon optimized):NOSt pICH47742

    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 Selectable Marker PI Publication Hidden Extra Search Info
    pBUN501zCas9D10A (Synthetic), gRNA scaffold (Synthetic)Ubi1p, AtU6-26pBar Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses zCas9D10A, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Bar resistance pGreen-like binary vector
    pHSN501zCas9D10A (Synthetic), gRNA scaffold (Synthetic)2×35Sp, AtU6-26pHygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses zCas9D10A, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Hyg resistance pGreen-like binary vector
    pDe-CAS9-D10ACas9-D10A (Arabidopsis thaliana)PcUbi4-2Kanamycin Puchta Both CRISPR/Cas-based nucleases and nickases can be used efficiently for genome engineering in Arabidopsis thaliana. Plant J. 2014 Jul;79(2):348-59. doi: 10.1111/tpj.12554. Epub 2014 Jun 17. based on pDe-CAS9, encodes Cas9 D10A nickase variant pPZP221
    pYPQ159hSpCas9D10A (Human codon optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with hSpCas9D10A pNJB185-linker6
    pDGE76pnos:nptII-tmos (Other), p2x35S:Cas9 D10A-tnos (Other), BsaI-ccdB_CmR-BsaI Stuttmann Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319. recipient plasmid [multiplexing, nickase]; p35S:Cas9 D10A, nptII pVM_BGW
    pDGE77pnos:PAT-tnos (Other), p2x35S:Cas9 D10A-tnos (Other), BsaI-ccdB_CmR-BsaI Stuttmann Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319. recipient plasmid [multiplexing, nickase]; p35S:Cas9 D10A, BASTA pVM_BGW
    pDGE78pnos:nptII-tmos (Other), pPcUbi:Cas9 D10A-tnos (Other), BsaI-ccdB_CmR-BsaI Stuttmann Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319. recipient plasmid [multiplexing, nickase]; pPcUbi:Cas9 D10A, nptII pVM_BGW
    pDGE79pnos:PAT-tnos (Other), pPcUbi:Cas9 D10A-tnos (Other), BsaI-ccdB_CmR-BsaI Stuttmann Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. Plant J. 2016 Aug 31. doi: 10.1111/tpj.13319. recipient plasmid [multiplexing, nickase]; pPcUbi:Cas9 D10A, BASTA pVM_BGW
    pTX179Hygromycin Voytas A Single Transcript CRISPR-Cas9 System for Efficient Genome Editing in Plants. Mol Plant. 2016 Jul 6;9(7):1088-91. doi: 10.1016/j.molp.2016.05.001. Epub 2016 May 19. Express STU (Single Transcriptional Unit) Cas9 nickase with Maize Ubiquitin1 promoter in plants pTX172
  • Tag / Fusion Protein
    • SV40 NLS (C terminal on insert)
    		•
    pMOD_A0201 AtCas9_D10A nickase (Synthetic) 35S Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: 35S, Gene: AtCas9_D10A nickase, Terminator: HSP pMOD_A1001
    pMOD_A0202 AtCas9_D10A nickase (Synthetic) AtUbi10 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: AtUbi10, Gene: AtCas9_D10A nickase, Terminator: HSP pMOD_A1001
    pMOD_A0208 AtCas9_D10A nickase (Synthetic) EC1.2 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: EC1.2, Gene: AtCas9_D10A nickase, Terminator: HSP pMOD_A1001
    pMOD_A0301 AtCas9_H840A nickase (Synthetic) 35S Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: 35S, Gene: AtCas9_H840A nickase, Terminator: HSP pMOD_A1001
    pMOD_A0302 AtCas9_H840A nickase (Synthetic) AtUbi10 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: AtUbi10, Gene: AtCas9_H840A nickase, Terminator: HSP pMOD_A1001
    pMOD_A0308 AtCas9_H840A nickase (Synthetic) EC1.2 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: EC1.2, Gene: AtCas9_H840A nickase, Terminator: HSP pMOD_A1001
    pMOD_A0601 Csy4-P2A-AtCas9_D10A nickase (Synthetic) 35S Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: 35S, Gene: Csy4-P2A-AtCas9_D10A nickase, Terminator: HSP pMOD_A1001
    pMOD_A0608 Csy4-P2A-AtCas9_D10A nickase (Synthetic) EC1.2 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: EC1.2, Gene: Csy4-P2A-AtCas9_D10A nickase, Terminator: HSP pMOD_A1001
    pMOD_A0701 Csy4-P2A-AtCas9_H840A nickase (Synthetic) 35S Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: 35S, Gene: Csy4-P2A-AtCas9_H840A nickase, Terminator: HSP pMOD_A1001
    pMOD_A0708 Csy4-P2A-AtCas9_H840A nickase (Synthetic) EC1.2 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: EC1.2, Gene: Csy4-P2A-AtCas9_H840A nickase, Terminator: HSP pMOD_A1001
    pMOD_A1210 TaCas9_D10A nickase (Synthetic) ZmUbi Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: ZmUbi, Gene: TaCas9_D10A nickase, Terminator: HSP pMOD_A1001
    pMOD_A1310 TaCas9_H840A nickase (Synthetic) ZmUbi Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: ZmUbi, Gene: TaCas9_H840A nickase, Terminator: HSP pMOD_A1001
    pMOD_A1610 Csy4-P2A-TaCas9_D10A nickase (Synthetic) ZmUbi Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: ZmUbi, Gene: Csy4-P2A-TaCas9_D10A nickase, Terminator: HSP pMOD_A1001
    pMOD_A1710 Csy4-P2A-TaCas9_H840A nickase (Synthetic) ZmUbi Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: ZmUbi, Gene: Csy4-P2A-TaCas9_H840A nickase, Terminator: HSP pMOD_A1001
    pAtCas9-D10A_1 AtCas9_D10A (nickase) (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9_D10A (nickase), Plant Selection: 2x35S:hpt II pCAMBIA
    pAtCas9-D10A_2 AtCas9_D10A (nickase) (Synthetic)Kanamycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9_D10A (nickase), Plant Selection: 2x35S:npt II pCAMBIA
    pAtCas9-D10A_3 AtCas9_D10A (nickase) (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9_D10A (nickase), Plant Selection: 2x35S:bar pCAMBIA
    pAtCas9-H840A_1 AtCas9_H840A (nickase) (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9_H840A (nickase), Plant Selection: 2x35S:hpt II pCAMBIA
    pAtCas9-H840A_2 AtCas9_H840A (nickase) (Synthetic)Kanamycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9_H840A (nickase), Plant Selection: 2x35S:npt II pCAMBIA
    pTaCas9-D10A_5 TaCas9_D10A (nickase) (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: TaCas9_D10A (nickase), Plant Selection: PvUbi2:hpt II pCAMBIA
    pTaCas9-D10A_6 TaCas9_D10A (nickase) (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: TaCas9_D10A (nickase), Plant Selection: PvUbi2:bar pCAMBIA
    pTaCas9-H840A_5 TaCas9_H840A (nickase) (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: TaCas9_H840A (nickase), Plant Selection: PvUbi2:hpt II pCAMBIA
    pTaCas9-H840A_6 TaCas9_H840A (nickase) (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: TaCas9_H840A (nickase), Plant Selection: PvUbi2:bar pCAMBIA
    pJG363nAtCas9_D10A+gNt_R2+donor (Other) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. BeYDV replicon T-DNA for gene targeting in tobacco leaves, D10A single nickase (nAtCas9_D10A+gNt_R2+donor) pTRANS_201
    pJG367nAtCas9_H840A+gNt_R2+gNt_F2+donor (Other) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. BeYDV replicon T-DNA for gene targeting in tobacco leaves, H840A double nickase (nAtCas9_H840A+gNt_R2+gNt_F2+donor) pTRANS_201
    pJG370nAtCas9_D10A+gRNActrl+donor (Synthetic) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. BeYDV replicon T-DNA for gene targeting in tobacco leaves, D10A single nickase no gRNA control (nAtCas9_D10A+gRNActrl+donor) pTRANS_201
    pJG382nAtCas9_D10A+gNt_R2+gNt_F2+donor (Other) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. BeYDV replicon T-DNA for gene targeting in tobacco leaves, D10A double nickase (nAtCas9_D10A+gNt_R2+gNt_F2+donor) pTRANS_201
    pJG484nTaCas9_H840A+gUbi8+donor (Other) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. WDV replicon T-DNA for gene targeting in wheat scutella, H840A single nickase (nTaCas9_H840A+gUbi8+donor) pTRANS_203
    pJG486nTaCas9_H840A+gUbi1+donor (Other) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. WDV replicon T-DNA for gene targeting in wheat scutella, H840A single nickase (nTaCas9_H840A+gUbi1+donor) pTRANS_203
    pJG488nTaCas9_H840A+gUbi8+gUbi1+donor (Other) Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. WDV replicon T-DNA for gene targeting in wheat scutella, H840A double nickase (nTaCas9_H840A+gUbi8+gUbi1+donor) pTRANS_203
    pORFE0002AtCAS9 nickase D10A (Other) Chevalier Chevalier Lab plasmids (unpublished) AtCAS9 nickase D10A module CDS1 pICH41308
    pORFE0014AtCAS9 D10A without stop codon (Other) Chevalier Chevalier Lab plasmids (unpublished) AtCAS9 nickase D10A module CDS1ns pAGM1287
    pORFE1052AtCAS9 D10A (Other)2x35S Chevalier Chevalier Lab plasmids (unpublished) Overexpression of the AtCAS9 nickase in plant under 2x35S promoter pICH47742
    pFH91nScCas9 (nickase), wheat codon optimized (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level0 nScCas9 (nickase), wheat codon optimized pICH41308
    pFH109ZmUBIp::nScCas9 (wheat codon optimized):NOSt (Synthetic) Nekrasov A modular cloning toolkit for genome editing in plants bioRxiv Level1 ZmUBIp::nScCas9 (wheat codon optimized):NOSt pICH47742

    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 Selectable Marker PI Publication Hidden Extra Search Info
    pBUN6A11dCas9-VP64 (Synthetic), gRNA scaffold (Synthetic)Ubi1p, OsU3pBar Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses dCas9-VP64, gRNA scaffold for insertion of target sequence (OsU3 promoter), Bar resistance pGreen-like binary vector
    pHSN6A01dCas9-VP64 (Synthetic), gRNA scaffold (Synthetic)2×35Sp, AtU6-26pHygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses dCas9-VP64, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Hyg resistance pGreen-like binary vector
    pYPQ152pco-dCas9-VP64 (Plant codon-optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with pco-dCas9-VP64 pYPQ185-linker1
    pYPQ173pco-dCas9-VP64-T2A-MS2-VP64 (Synthetic) Qi Robust transcriptional activation in plants using multiplexed CRISPR-Act2.0 and mTALE-Act systems. Mol Plant. 2017 Nov 29. pii: S1674-2052(17)30344-1. doi: 10.1016/j.molp.2017.11.010. Express CRISPR-Act2.0 system which contains pco-dCas9-VP64 fusion protein and MS2-VP64 fusion protein linked by in-frame T2A (encoding Thosea asigna 'self-cleaving' 2A peptide) sequence unknown
    PV225dCas9-AT-CBF1 (Other) Young Repurposing of Type I-E CRISPR-Cascade for gene activation in plants Nature Communications Biology dCas9 plant transcriptional activator (AT-CBF1) linked constitutive expression cassette for Zea mays PHP17720
  • Tag / Fusion Protein
    • AT-CBF1 (C terminal on insert)
    • CBF1 AT4G25490, ATCBF1, C-repeat/DRE binding factor 1, DRE BINDING PROTEIN 1B, DREB1B, T30C3.11, TRANSCRIPTIONAL ACTIVATOR CBF1

    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 Selectable Marker PI Publication Hidden Extra Search Info
    pBUN6I11dCas9-KRAB (Synthetic), gRNA scaffold (Synthetic)Ubi1p, OsU3pBar Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses dCas9-KRAB, gRNA scaffold for insertion of target sequence (OsU3 promoter), Bar resistance pGreen-like binary vector
    pHSN6I01dCas9-KRAB (Synthetic), gRNA scaffold (Synthetic)2×35Sp, AtU6-26pHygromycin Chen A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 2014 Nov 29;14(1):327. CRISPR/Cas based plant genome editing and gene regulation; expresses dCas9-KRAB, gRNA scaffold for insertion of target sequence (AtU6-26 promoter), Hyg resistance pGreen-like binary vector
    pEGB 35S:dCas9:Tnos (GB1191)dCas9 (Other)35S Orzaez GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. Plant Physiol. 2013 Jul;162(3):1618-31. Transcriptional unit for human codon optimized with mutated (D10A, H840A) and inactivated catalytic domains Cas9 protein plant expression driven by the 35S promoter pDGB3alpha2
    pYPQ153pco-dCas9-3X(SRDX) (Plant codon-optimized) (Other) Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 2015 Aug 21. pii: pp.00636.2015. Gateway entry vector with pco-dCas9-3X(SRDX) pYPQ185-linker1
    pdCas9 (GB1079)Cas9 coding region with mutated (D10A, H840A) and inactivated catalytic domains (human codon optimised) (Other) Orzaez A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. eCollection 2016. Provides the human codon optimized CDS of Cas9 protein with mutated (D10A, H840A) and inactivated catalytic domains as a level 0 GoldenBraid part for C-terminal fusions pUPD
    pEGB 35s:dCas:BRD:tNos (GB1172)dCas9:BRD (Synthetic)35S Orzaez A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. eCollection 2016. Transcriptional unit of (human codon optimized) inactivated Cas9 fused to the BRD Transcriptional Repressor pDGB3alpha2
    pYPQ223 (dAsCpf1-SRDX)dAsCpf1-SRDX (Other) Qi A CRISPR-Cpf1 system for efficient genome editing and transcriptional repression in plants. Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18. Deactivated AsCpf1-SRDX repressor fusion; transcriptional repressor dCpf1 Gateway entry plasmid pYPQ221
    pYPQ233 (dLbCpf1-SRDX)LbCpf1-SRDX (Other) Qi A CRISPR-Cpf1 system for efficient genome editing and transcriptional repression in plants. Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18. Deactivated LbCpf1-SRDX repressor fusion; transcriptional repressor dCpf1 Gateway entry plasmid pYPQ221
    pMOD_A0402 AtCas9_dead (D10A + H840A) (Synthetic) AtUbi10 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: AtUbi10, Gene: AtCas9_dead (D10A + H840A), Terminator: HSP pMOD_A1001
    pMOD_A0408 AtCas9_dead (D10A + H840A) (Synthetic) EC1.2 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: EC1.2, Gene: AtCas9_dead (D10A + H840A), Terminator: HSP pMOD_A1001
    pMOD_A0801 Csy4-P2A-AtCas9_dead (D10A + H840A) (Synthetic) 35S Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: 35S, Gene: Csy4-P2A-AtCas9_dead (D10A + H840A), Terminator: HSP pMOD_A1001
    pMOD_A0802 Csy4-P2A-AtCas9_dead (D10A + H840A) (Synthetic) AtUbi10 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: AtUbi10, Gene: Csy4-P2A-AtCas9_dead (D10A + H840A), Terminator: HSP pMOD_A1001
    pMOD_A0808 Csy4-P2A-AtCas9_dead (D10A + H840A) (Synthetic) EC1.2 Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: EC1.2, Gene: Csy4-P2A-AtCas9_dead (D10A + H840A), Terminator: HSP pMOD_A1001
    pMOD_A1410 TaCas9_dead (D10A + H840A) (Synthetic) ZmUbi Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: ZmUbi, Gene: TaCas9_dead (D10A + H840A), Terminator: HSP pMOD_A1001
    pMOD_A1810 Csy4-P2A-TaCas9_dead (D10A + H840A) (Synthetic) ZmUbi Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Module A, Promoter: ZmUbi, Gene: Csy4-P2A-TaCas9_dead (D10A + H840A), Terminator: HSP pMOD_A1001
    pDIRECT_21B Engineering Reagent: 35S:AtCas9_dead + AtU6:gRNA (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: 35S:AtCas9_dead + AtU6:gRNA, Plant Selection: 2x35S:hpt II pTRANS_210
    pDIRECT_25G Engineering Reagent: ZmUbi:TaCas9_dead + TaU6:gRNA (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: ZmUbi:TaCas9_dead + TaU6:gRNA , Plant Selection: PvUbi2:hpt II pTRANS_250d
    pDIRECT_26G Engineering Reagent: ZmUbi:TaCas9_dead + TaU6:gRNA (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Direct Cloning, Type: T-DNA, Engineering Reagent: ZmUbi:TaCas9_dead + TaU6:gRNA , Plant Selection: PvUbi2:bar pTRANS_260d
    pAtCas9-dead_1 AtCas9_D10A+H840A (dead) (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9_D10A+H840A (dead), Plant Selection: 2x35S:hpt II pCAMBIA
    pAtCas9-dead_2 AtCas9_D10A+H840A (dead) (Synthetic)Kanamycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9_D10A+H840A (dead), Plant Selection: 2x35S:npt II pCAMBIA
    pAtCas9-dead_3 AtCas9_D10A+H840A (dead) (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: AtCas9_D10A+H840A (dead), Plant Selection: 2x35S:bar pCAMBIA
    pTaCas9-dead_5 TaCas9_D10A+H840A (dead) (Synthetic)Hygromycin Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: TaCas9_D10A+H840A (dead), Plant Selection: PvUbi2:hpt II pCAMBIA
    pTaCas9-dead_6 TaCas9_D10A+H840A (dead) (Synthetic)Basta Voytas A multi-purpose toolkit to enable advanced genome engineering in plants. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. Cas9 only expression, Cas9 Type: TaCas9_D10A+H840A (dead), Plant Selection: PvUbi2:bar pCAMBIA
    pHdzCas9-KRABCaMV35SHygromycin Ng CRISPRi mediated phosphoenolpyruvate carboxylase regulation to enhance the production of lipid in Chlamydomonas reinhardtii. Bioresour Technol. 2017 May 4. pii: S0960-8524(17)30619-3. doi: 10.1016/j.biortech.2017.04.111. Expression dzCas9-KRAB in microalgae. CRISPR/Cas based plant genome editing and gene regulation, Hyg resistance pHdzCas9-KRAB
  • Tag / Fusion Protein
    • KRAB (C terminal on insert)
    		•
    pORFE0003AtDeadCAS9 D10A H840A (Other) Chevalier Chevalier Lab plasmids (unpublished) AtDeadCAS9 D10A H840A module CDS1 pICH41308
    pORFE0005AtDeadCAS9 D10A H840A without stop codon (Other) Chevalier Chevalier Lab plasmids (unpublished) AtDeadCAS9 D10A H840A without stop codon module CDS1ns pAGM1287

    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.

    Plasmid Promoter Cloning Enzyme Co-expressed Cas9 Cas9 System Selection PI
    pICSL01009::AtU6p aU6 BsaI none S. pyogenes Kamoun
    pUC119-gRNA U6 PCR template none S. pyogenes Sheen
    pRGEB31 rice snoRNA U3 BsaI none S. pyogenes Yang
    pBUN6A11 OsU3 BsaI yes, activate S. pyogenes Bar Chen
    pBUN6I11 OsU3 BsaI yes, interfere S. pyogenes Bar Chen
    pBUN501 AtU6-26 BsaI yes, nick S. pyogenes Bar Chen
    pCBC-MT2T3 see paper BsaI none S. pyogenes Chen
    pCBC-MT3T4 see paper BsaI none S. pyogenes Chen
    pBUN421 TaU3 BsaI yes, cut S. pyogenes Bar Chen
    pU3-gRNA OsU3 AarI none S. pyogenes Gao
    pZmU3-gRNA maize U3 none S. pyogenes Gao
    pU6-gRNA wheat U6 BbsI none S. pyogenes Gao
    pBlu/gRNA Arabidopsis U6 BbsI none S. pyogenes Stupar
    pBUE411 OsU3 yes, cut S. pyogenes Basta Chen
    pHUE411 OsU3 yes, cut S. pyogenes Hyg Chen
    pHAtC U6 AarI yes, cut S. pyogenes Hygro Kim
    pBAtC U6 AarI yes, cut S. pyogenes Basta Kim
    pHEE401 U6-26p BsaI yes, cut S. pyogenes Hygro Chen
    pHEE401E U6-26p BsaI yes, cut S. pyogenes Hygro Chen
    pHDE-35S-Cas9-mCherry-UBQ U6-26 PmeI yes, cut S. pyogenes Hygro, mCherry Zhao
    pCBC-DT1T1 see paper BsaI none S. pyogenes Chen
    pRGEB32 rice snoRNA U3 BsaI yes, cut S. pyogenes Hygro Yang
    pHSE401 AtU6-26 yes, cut S. pyogenes Hyg Chen
    pBUN411 OsU3 BsaI yes, cut S. pyogenes Bar Chen
    pHSN6A01 AtU6-26 BsaI yes, activate S. pyogenes Hyg Chen
    pHSN6I01 AtU6-26 BsaI yes, interfere S. pyogenes Hyg Chen
    pHSN501 AtU6-26 BsaI yes, nick S. pyogenes Hyg Chen
    pCBC-DT3T4 see paper BsaI none S. pyogenes Chen
    pCBC-MT1T2 see paper BsaI none S. pyogenes Chen
    pRGE32 rice snoRNA U3 BsaI yes, cut S. pyogenes Hygro Yang
    pKSE401 AtU6-26 yes, cut S. pyogenes Kan Chen
    pRGE31 rice snoRNA U3 BsaI none S. pyogenes Yang
    pUC gRNA Shuttle Mt U6.6 inFusion none S. pyogenes Parrott
    pCBC-DT2T3 see paper BsaI none S. pyogenes Chen
    pHSN401 AtU6-26 BsaI yes, cut S. pyogenes Hyg Chen
    pYPQ141-ZmUbi-RZ-As Maize ubiquitin 1 none As Cpf1 Qi
    pKEE401 yes, cut S. pyogenes Neomycin Chen

    Do you have suggestions for other plasmids that should be added to this list?

    Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource!