...described here. #60224 - AAV:ITR-U6-sgRNA(Kras)-U6-sgRNA(p53)-U6-sgRNA(Lkb1)-pEFS-Rluc-2A-Cre-shortPA-KrasG12D_HDRdonor-ITR...lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9-Blast and lentiGuide-Puro (sgRNA) Browse...CMV-driven SaCas9; U6-driven sgRNA 61593 : PX602; TBG-driven SaCas9; U6-driven sgRNA 61592 : PX600; CMV-driven... SaCas9 sgRNA can be co-transfected as PCR amplicons using a protocol similar to SpCas9 sgRNA PCR (Ran... pAAV-U6::sgRNA(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). This is a AAV plasmid for sgRNA cloning. GFP-KASH...implemented by delivering Cre and sgRNA to a Cre-dependent mouse or sgRNA to a constitutively Cas9-expressing...control for AAV-KPL; sgRNA targeted to LacZ, plus luciferase-2A-Cre recombinase 60226 : sgRNA cloning backbone...

...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...targeting 19,114 genes and containing 76,441 unique sgRNAs along with 1000 non-targeting controls. This backbone...backbone does not contain SpCas9. 76,441 unique sgRNAs targeting 19,114 human genes along with 1000 non-targeting...Human gRNA pooled library in lentiCRISPRv2 Human sgRNA library in backbone lentiCRISPRv2 targeting 19,114...

...dCas9) protein and a customized s ingle g uide RNA (sgRNA) designed with a 20-basepair complementary region...any gene of interest. Co-expression of dCas9 and sgRNA can efficiently block transcription (in bacteria...determined jointly by 20-bp matching region on the sgRNA and a short DNA motif (protospacer adjacent motif...plasmid ( pgRNA_bacteria ) contains a constitutive sgRNA expression cassette on a ColE1 vector with ampicillin...pgRNA_humanized ) contains a murine U6 promoter controlled sgRNA cassette, wherein the GN19 can be custom designed...pSLQ1651-sgTelomere(F+E) Lentiviral vector with sgRNA targeting human telomeres 46913 pMSCV-LTR-dCas9-...pSico-based U6 vector containing murine U6 promoter and sgRNA targeting GFP (NT1) 46917 pU6-sgCXCR4-2 Human pSico-based...

...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... via the links below: Table 1. dCas9 Activators sgRNA Dual Expression ID Plasmid 48236 pAC2-dual-dCas9VP48...expression construct expressing both dCas9VP48 and sgRNA from separate promoters. 48238 pAC152-dual-dCas9VP64...expression construct expressing both dCas9VP64 and sgRNA from separate promoters. 48239 pAC153-dual-dCas9VP96...expression construct expressing both dCas9VP64 and sgRNA from separate promoters. 48240 pAC154-dual-dCas9VP160...

... nicking sgRNA BbsI No Vermilion Norbert Perrimon pHSG1C3 Mammalian U6 pegRNA + nicking sgRNA BbsI for...Liu U6-pegRNA-H1-nick sgRNA-mCherry Mammalian, AAV hU6 + H1 pegRNA + nicking sgRNA No mCherry Hyongbum ... HindIII No Tilmann Weber pnsgRNA Bacteria J23119 (BBa_J23119) nicking sgRNA No Tilmann Weber pPBT-peRNA_GG-Puro...fusion enzyme and pegRNA plus an additional simple sgRNA, which directs the Cas9 nickase to nick the unedited...increases indel formation slightly. Designing the sgRNA with a spacer that only binds the edited strand,... the PE2 enzyme. In the PE3 and PE3b systems an sgRNA targeting the opposite strand is added . Browse,...the type of gRNA (e.g., pegRNA, epegRNA, nicking sgRNA), cloning enzyme, selectable marker, and whether...

...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...dCas9-VP64 along with 1-4 sgRNAs expressed from independent promoters. sgRNAs are cloned into separate ... and allow for expression of more sgRNAs in a single vector. sgRNAs are expressed as polycistronic glycine...pLenti-sgRNA-Lib 53121 Mammalian/Lentiviral BsmBI none S. pyogenes ccdB Wei pLKO5.sgRNA.EFS.PAC 57825 ...Church LRG (Lenti_sgRNA_EFS_GFP) 65656 Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA(F+E) 59986... Boxem PU6::klp-12_sgRNA 46170 Worm PCR (see paper) none S. pyogenes Calarco SP6-sgRNA-scaffold 47912 ...

...example, sgRNA-A’s protospacer sequence and PAM happen to fall on the bottom (Crick) strand while sgRNA-B’s...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...non-coding elements. For creating a gene knockout, two sgRNA located within exons will enrich even monoallelic...

...S. pyogenes sgRNA from the H1 promoter. 53187 pmU6-gRNA : Expresses the S. pyogenes sgRNA from the mouse...pyogenes sgRNA from the human U6 promoter. 53189 ph7SK-gRNA : Expresses the S. pyogenes sgRNA from the...directed to its target in gene promoters by the sgRNA expressed in trans from pSPgRNA, the dCas9-VP64 ...activates the expression of downstream genes. Multiple sgRNAs targeted to the same promoter act synergistically...

... Sabatini and Lander gRNA cloning into pLX-sgRNA pLX-sgRNA PDF 125.9 KB Sontheimer gRNA design and cloning...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 ...Cas9 target online predictor Identifies candidate sgRNA target sites by off-target quality. Validated for...Goldstein Nematode: gRNA design and cloning Cas9-sgRNA construct PDF 355.3 KB Goldstein Nematode: Injection...

...PU6::unc-119_sgRNA : unc-119targeting sgRNA. 46170 PU6::klp-12_sgRNA : klp-12targeting sgRNA. Please note.... elegans . The protocol for customizing the sgRNA expression plasmid can be located in the Materials...

...46,354 BARBEKO sgRNA Library 174163 Knockout, Base Editing Human Wei 3rd 3 53,502 sgRNA Pool for Streptococcus...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,... Mouse Knouse 3rd ~5 71,878 Mouse messenger-RBP sgRNA Library 169082 Knockout Mouse Turner N/A 10 13,500...

... an sgRNA uder the Arabidopsis U6 promoter. 46966 pICH86966::AtU6p::sgRNA_PDS : Expresses an sgRNA targeting...manuscript for the protocol on assembling customised sgRNA expressing plasmids. Questions regarding the protocol...module. 49772 pAGM4723::AtU6p::sgRNA2-2x35S-5'UTR::Cas9::NOST-AtU6p::sgRNA1 : Level 2 construct....

...expression, human optimized 48673 M-NM-sgRNA Mammalian U6-driven sgRNA (NMm1) targeting GTCCCCTCCACCCCACAGTG...expression, human optimized 48672 M-ST1-sgRNA Mammalian U6-driven sgRNA (STm1) targeting GTCCCCTCCACCCCACAGTG...III at five protospacers 48671 M-SP-sgRNA Mammalian U6-driven sgRNA (SPm) targeting GTCCCCTCCACCCCACAGTG...

...PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid Calarco SP6-sgRNA-scaffold ...pyogenes (PAM = NGG) pDD162 (Peft-3::Cas9 + Empty sgRNA) R07E5.16 U6 yes, cut Goldstein DR274 T7 BsaI In...none, need Cas9 plasmid Sternberg L4440_BioBrick-sgRNA T7 BbsI, BsaI Ingestion by C. elegans none, need...

...vectors for the delivery of the synthetic guide RNA (sgRNA). Each of the Cas9 vectors contains the Cas9 coding...with and without a C-terminal EGFP fusion. For the sgRNA, vector pMB60 allows in vitro production of RNA ...

...60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc-2A-Cre-WPRE-hGHpA-ITR Cre, luciferase, and sgRNA expression EFS... Zhang 60229 AAV:ITR-U6-sgRNA(backbone)-pCBh-Cre-WPRE-hGHpA-ITR Cre and sgRNA coexpression Cbh AAV Zhang...ITR-U6-sgRNA(backbone)-hSyn-Cre-2A-EGFP-KASH-WPRE-shortPA-ITR Cre, KASH-tagged EGFP, and sgRNA expression...expression hSyn AAV Zhang 60820 pSECC Cre, Cas9 and sgRNA expression EF-1 alpha Lentiviral Jacks 60877 pMAZe...

... plate, co-deliver into the cells with the Cas9/sgRNAs vector (like PX330). For large changes (>100bp ... a typical size (~ 5kb) donor plasmid with Cas9/sgRNAs vector, for one well in 24-well plate. A good example...of a typical size (~5kb) donor plasmid with Cas9/sgRNAs vector, for a one well in a 24-well plate. A good...

...Cloning Vector Bbs I ver. 2 An empty sgRNA expression vector. sgRNA sequence can be cloned into BbsI site...

...replacement by homologous recombination. For the sgRNA, vector pMB60 allows in vitro production of RNA ...

...Virmilion none, need Cas9 plasmid Bullock and Port pAc-sgRNA-Cas9 dU6 BspQI Transfection Puromycin yes, cut Ji-Long...