... in two formats: 1 vector system - lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9...described here. #60224 - AAV:ITR-U6-sgRNA(Kras)-U6-sgRNA(p53)-U6-sgRNA(Lkb1)-pEFS-Rluc-2A-Cre-shortPA-KrasG12D_HDRdonor-ITR...below) can be cloned scarlessly into the vector before the sgRNA scaffold. The oligos are designed based...lentiviral vector encoding dCAS9-VP64 with 2A Blast resistance marker 61426 : lentiviral vector encoding... both single- and dual-vector systems that allow the delivery of Cas9 and sgRNA in vivo via adeno-associated...single guide RNA (sgRNA) into one plasmid (Ran et al., Nature 2015). The dual vector system uses S. pyogenes...SpCas9), using one vector to express SpCas9, and another to express up to three sgRNAs (Swiech et al., Nature...

...more details about our viral vector packaging service, visit our Viral Vector Packaging Service page . Pooled...pooled libraries 73178 Human sgRNA library Brunello in lentiGuide-Puro Human sgRNA library in backbone lentiGuide-Puro...Human CRISPRa sgRNA library Calabrese in backbone XPR_502 (P65 HSF), Set A Human CRISPRa sgRNA activation...containing 56,762 unique sgRNAs. This backbone does not contain dCas9. 56,762 unique sgRNAs targeting 18,885 ...pooled libraries 73633 Mouse sgRNA library Brie in lentiGuide-Puro Mouse sgRNA library in backbone lentiGuide-Puro...Ready-to-use lentivirus available from Addgene's viral vector packaging service. Browse constructs currently ... Viral Vector Packaging Service Lentivirus Browse In-Stock Lentivirus...

...pgRNA_bacteria ) contains a constitutive sgRNA expression cassette on a ColE1 vector with ampicillin resistance, wherein...51024 pSLQ1651-sgTelomere(F+E) Lentiviral vector with sgRNA targeting human telomeres 46913 pMSCV-LTR-...sgGFP-NT1 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GFP (NT1) 46917 pU6...sgCXCR4-2 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting endogenous CXCR4 .../ARS vector (Leu2) that contains dCas9 fused to N 51025 pSLQ1661-sgMUC4-E3(F+E) Lentiviral vector that...pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GAL4 UAS promoter 46916 pU6-sgGAL4-4...-4 Human pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GAL4 UAS promoter (negative...

...guide RNA (sgRNA). While the dCas9 protein provides the transactivation activity, the sgRNA component ...activators and sgRNAs can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA expression...expression vector or the linear DNA containing the sgRNA expressing fragment produced by PCR), or expressed from...Gateway donor vectors [48218~48221] and can be transferred to Gateway destination vectors (e.g., pmax-DEST...pmax expression vector. 48224 pAC92-pmax-dCas9VP96 : dCas9VP96 on pmax expression vector. 48225 pAC93-pmax-dCas9VP160...Gateway donor vector pCR8/GW/TOPO. 48218 pAC84-pCR8-dCas9 : dCas9 on Gateway donor vector pCR8/GW/TOPO....Gateway donor vector pCR8/GW/TOPO. 48220 pAC148-pCR8-dCas9VP96 : dCas9VP96 on Gateway donor vector pCR8/GW/...

... Empty Vectors Many gRNA empty vectors have been deposited at Addgene. To find the gRNA vector for your... and allow for expression of more sgRNAs in a single vector. sgRNAs are expressed as polycistronic glycine...Mammalian BbsI none S. pyogenes Gersbach pUC57-sgRNA expression vector 51132 Mammalian BsaI none S. pyogenes Huang...pyogenes Joung sgRNA with U6 promoter 48962 Worm HindIII none S. pyogenes de Bono sgRNA with rpr-1 promoter...SpBsmBI sgRNA Hygro 62205 Mammalian/Lentiviral BsmBI none S. pyogenes Hygro Maehr pLenti SpBsmBI sgRNA Puro...Bacteria T7 yes, cut N. meningitidis Joung Cas9 sgRNA vector 68463 Mammalian U6 none S. pyogenes Zhang Cas9...Multiplex gRNA Vectors Several systems have been developed to generate multiplex gRNA vectors. These systems...

...Publication Empty Prime Editing gRNA Vectors A selection of empty gRNA vectors suitable for prime editing are...nicking sgRNA BbsI No Vermilion Norbert Perrimon 164423 pHSG1C3 Mammalian U6 pegRNA + nicking sgRNA BbsI ... RNA-H1-nick sgRNA-mCherry Mammalian, AAV hU6 + H1 pegRNA + nicking sgRNA No mCherry Hyongbum ...HindIII No Tilmann Weber 172717 pnsgRNA Bacteria J23119 (BBa_J23119) nicking sgRNA No Tilmann Weber 173220 pPBT-peRNA_GG-Puro...-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila ...mutations in RT enzyme PE3 - PE2 plus additional sgRNA PE4 - PE2 plus MLH1 (a protein component of MMR)... table below. Use the search bar to find a gRNA vector based on expression system, promoter, the type ...

... gRNA design and cloning gRNA cloning vector Retroviral vectors: neomycin (pSIR-neo) , GFP (pSIR-GFP) ... Sabatini and Lander gRNA cloning into pLX-sgRNA pLX-sgRNA PDF 125.9 KB Sontheimer gRNA design and cloning... pooled library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9-Blast and lentiGuide-Puro...as an optional input to assess HDR frequency. An sgRNA sequence (without PAM sequence) can be provided,... can identify positively or negatively selected sgRNAs, genes, or pathways. CrispyCrunch CrispyCrunch ...CRISPR in 180+ genomes. PubMed PMID: 27380939 . sgRNA Scorer 2.0 From the Church Lab : a tool that identifies... epression, and a ctivation From the Qi Lab , a sgRNA design tool for genome editing, as well as gene ...

...example, sgRNA-A’s protospacer sequence and PAM happen to fall on the bottom (Crick) strand while sgRNA-B’s... the 20-mer is not G. sgRNA expression from the U6 promoter of the pX330 vector is enhanced by the inclusion...primers flanking each sgRNA target site (within 150 - 350 bp) to amplify the sgRNA target site to examine...complement: 1.0 μl sgRNA 24- or 25-mer oligo (100 μM; see step 1.4 ), 1.0 μl sgRNA 24- or 25-mer reverse...composed of a polyclonal population exposed to sgRNA-A and sgRNA-B (see step 5 ). The purpose of the following...delivery of a pair of chimeric single guide RNAs (sgRNAs) to create two double strand breaks (DSBs) at a...posted at the top of the page. CRISPR Design Design sgRNAs manually or using freely available online tools...

...46,354 BARBEKO sgRNA Library 174163 Knockout, Base Editing Human Wei 3rd 3 53,502 sgRNA Pool for Streptococcus...-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila ...131625 Inhibition E. coli Bikard N/A ∼5 21,417 Bison sgRNA Library 169942 Knockout Human Ebert 3rd 4 2,852 ...Fly Perrimon 3rd 8 6 6 7,956 17,827 59,406 Dual sgRNA CRISPRi Libraries 187246 — 187248 1000000192 Inhibition...Inhibition Human Weissman 3rd 6 Varies Dual sgRNA CRISPRa Libraries 187249 — 187251 1000000193 Activation...3rd 2 37,722 Green monkey (Chlorocebus sabaeus) sgRNA library 178284 Knockout Green Monkey Doench and ...Knockout Human Parts 3rd 3 59,576 Human Whole Genome sgRNA iBAR Library 140633 Knockout Human Wei 3rd 3 58,...

...-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila ... Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...both the gRNA and Cas protein act as all-in-one vectors, but their function is often limited to a single... (PAM = NGG) 47549 pDD162 (Peft-3::Cas9 + Empty sgRNA) R07E5.16 U6 yes, cut Goldstein 42250 DR274 T7 BsaI...transcription none, need Cas9 plasmid Boxem 47912 SP6-sgRNA-scaffold SP6 AflII In vitro transcription none, ...need Cas9 plasmid Sternberg 177783 L4440_BioBrick-sgRNA T7 BbsI, BsaI Ingestion by C. elegans none, need...transcription none, need Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9...

...Cre-ERT2 - Tamoxifen inducible; Gateway entry vector none Entry Vector Zon 27493 pBT140 Cre CMV Mammalian Luo...Cre-ERT2;Targeting vector for Fos locus Mammalian Luo 46389 pArcCreER Cre-ERT2;Targeting vector for Arc locus...-ERT2; Targeting vector for Nanog locus mNanog Mammalian Hanna 60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc...2A-Cre-WPRE-hGHpA-ITR Cre, luciferase, and sgRNA expression EFS AAV Zhang 60229 AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-...pCBh-Cre-WPRE-hGHpA-ITR Cre and sgRNA coexpression Cbh AAV Zhang 60231 AAV:ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR...KASH-tagged EGFP, and sgRNA expression hSyn AAV Zhang 60820 pSECC Cre, Cas9 and sgRNA expression EF-1 alpha...Burgess 61570 Pvalb-2A-Cre targeting vector Cre; Targeting vector for Pvalb locus Mammalian Zeng 61573...

...plate, co-deliver into the cells with the Cas9/sgRNAs vector (like PX330). For large changes (>100bp insertions...typical size (~ 5kb) donor plasmid with Cas9/sgRNAs vector, for one well in 24-well plate. A good example...typical size (~5kb) donor plasmid with Cas9/sgRNAs vector, for a one well in a 24-well plate. A good reference...oligo when cloning my target spacers into the PX330 vector backbone? There is no need to add the NGG PAM sequence...Recombination (HR) FAQs Can you summarize HR template vector design guidelines? A few notes below are considerations...recently is the double nickase system, with PX335 vector, you need to find two guides for each cleavage ...

...interest 85586 gRNA Cloning Vector Bbs I ver. 2 An empty sgRNA expression vector. sgRNA sequence can be cloned...recognizing a telomere repeat. A retroviral expression vector with the puromycin resistance gene 63591 3xFN-Tel-TAL...recognizing a telomere repeat. A retroviral expression vector with the neomycin resistance gene 64325 3xFLAG-...

...replacement by homologous recombination. For the sgRNA, vector pMB60 allows in vitro production of RNA from...RNA polymerase III transcribed U6 snRNA. In both vectors, the target site sequence can be added by inserting...inserting an oligonucleotide linker into BsaI digested vector. Cloning targeting chiRNAs Targeting chiRNAs are...Drosophila snRNA:U6:96Ab promoter. 51019 pDsRed-attP : Vector for generating dsDNA donors for homology-directed...in combination with tracrRNA. 51434 pHD-DsRed : vector for generating dsDNA donors for homology-directed...

...-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila ... Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...both the gRNA and Cas protein act as all-in-one vectors, but their function is often limited to a single...none, need Cas9 plasmid Bullock and Port 49330 pAc-sgRNA-Cas9 dU6 BspQI Transfection Puromycin yes, cut Ji-Long...

...targeting vector assembly, and the remaining 5 plasmids include Cre and FLP expression vectors, a general...-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila ...Cas plasmid-system consists of two components: A vector containing Cas9 and a validated gRNA, based on ...the isolation of native protein complexes. This vector serves as a backbone to clone the left and right...Alternatively, cDNAs can be cloned directly into this vector and targeted to the AAVS1 genomic safe harbor locus...gRNA_AAVS1-T2 (Addgene #41818) or using an all in one vector from the Doyon lab, eSpCas9(1.1)_No_FLAG_AAVS1_...of HDR, which makes it easy to construct a donor vector, because MMEJ utilizes very short (around 20 bp...

...format into lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which Cas9 is included...systems contain two components: a guide RNA ( gRNA or sgRNA ) and a CRISPR-associated endonuclease (Cas enzyme...targeting in the double digits. These multiplex gRNA vectors can be used to knock out, activate, or repress ...multiplexing . Browse Plasmids: Multiplex gRNA Vectors Engineering the Cas9 Endonuclease CRISPR specificity...using a prime editing guide RNA ( pegRNA ). Like sgRNA, pegRNA directs the nickase to the target site by...efficiency PE3 - increased efficiency; an additional sgRNA directs Cas9n to a nearby site, driving mismatch...heterogeneous population of lentiviral transfer vectors. In most cases, each plasmid contains an individual...

...regions Joshua Mendell 212936 pGL3 Basic Vector Firefly Vector for investigating regions controlling transcription...interest are cloned into a Firefly luciferase reporter vector via Gateway cloning to test for the presence of...pGL4.84(hRlucCP/Puro) RapidResponse(TM) Renilla Vector for investigating regions controlling transcription...Pete Stecha 212933 pGL4.84(hRlucCP/Puro) Renilla Vector for investigating regions controlling transcription...Luciferase Type Promoter Description PI 60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Renilla...expression of firefly luciferase Paul Khavari 109395 EFS-Vector Firefly EF1α AAV-CRISPR knockout plasmid. Firefly...moniter viral transfection Sidi Chen 109396 TBG-Vector Firefly TBG AAV-CRISPR knockout plasmid. Firefly...

...Walker Jackson 78624 sgRNA(MS2)_Prnp_SAM1 PRNP U6 Dementia Walker Jackson 78625 sgRNA(MS2)_Prnp_SAM2 PRNP...19 James Trimmer 129409 Slc1a3-CreERT2 Targeting Vector SLC1A3 PGK Episodic ataxia Walker Jackson 129410...His T5 Parkinson's Suzanne Pfeffer 208392 Lenti-sgRNA-Cre-GpNLuc-sgmATM ATM Ataxia telangiectasia Gaorav...TARDBP-Exon1-ko-sgRNA1-SpCas9 TARDBP U6 ALS Rajat Rohatgi 107858 pHBS953 TARDBP-Exon2-ko-sgRNA2-SpCas9 TARDBP...133762 px459-TDP-43 sgRNA1 TARDBP U6 ALS Shawn Ferguson 133763 px459-TDP 43 sgRNA2 TARDBP U6 ALS Shawn...TIA1 His, T7 T7 ALS Thomas Tuschl 106108 px458_2A_GFP_sgRNA_EIF2AK2 EIF2AK2 U6 VWM disease Thomas Tuschl ...Episodic ataxia Xiaolong Wang 159788 pGL3-U6-sgRNA_KCNA1-mcherry KCNA1 U6 Episodic ataxia Xiaolong Wang...