...System Design gRNA Know your Cell Line Select Gene Select gRNA Clone gRNA Delivery of gRNA & Cas9 Validate...how to design your gRNA . Browse Plasmids: gRNAs Synthesize and clone desired gRNAs Once you have selected...enzyme and gRNA Species of promoter and expression pattern of promoter for Cas enzyme and gRNA Presence ...delivery of Cas9 and gRNA In vitro transcription reactions generate mature Cas9 mRNA and gRNA, which are then... Cas9-gRNA ribonucleoprotein (RNP) complexes Purified Cas9 protein and in vitro transcribed gRNA are combined...modify prior to designing your gRNA, as sequence variation between your gRNA targeting sequence and target...the associated gRNAs based on their predicted on-target and off-target activity ( see gRNA design software...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila... CRISPR gRNAs CRISPR Plasmids: gRNAs Browse CRISPR Plasmids...Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The table below lists gRNA plasmids designed to... questions to consider when choosing a gRNA: Does a given gRNA sequence exactly match your genomic target...Variation between a given gRNA sequence and your genomic target may reduce the gRNA activity. The variety ...pyogenes , S. aureus etc.) was this gRNA sequence designed for? A given gRNA sequence may only be compatible...gRNA expression vectors available from Addgene. CRISPR...

... protocol Church gRNA design and cloning gRNA cloning vector PDF 115.2 KB Church gRNA design and cloning...Nematode: Cas9 and gRNA use Cas9 (pIK86) ; gRNA empty backbone (pDR274) PDF 87.2 KB Liu Fly: gRNA cloning pAc-sgRNA-Cas9...Cas9 ; pUC gRNA Shuttle PDF 488.6 KB Sabatini and Lander gRNA cloning into pLX-sgRNA pLX-sgRNA PDF 125.9...the Chan Zuckerberg Biohub . gRNA Design Deep learning models to design gRNAs and predict their efficiency...Chen and Wente Zebrafish: gRNA cloning, in vitro transcription, injection gRNA core ; Cas9 ; optimized ...optimized Cas9 PDF 66.8 KB Fujii gRNA design and cloning gRNA cloning vector Retroviral vectors: neomycin (pSIR-neo...pluripotent stem cells pCas9_GFP ; gRNA empty vector Link O'Connor-Giles Fly: gRNA and ssODNs design and cloning...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...Selectable Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...

... the guide RNAs ( gRNAs ) used in these experiments are listed below. Note that gRNA produced from each... vector) 42241 DR299 (gRNA targeted to zebrafish gene apoea ) 42242 DR336 (gRNA targeted to zebrafish ...drd3 ) 42243 DR279 (gRNA targeted to zebrafish gene fh [site #1]) 42244 DR302 (gRNA targeted to zebrafish...site #2]) 42245 DR338 (gRNA targeted to zebrafish gene gsk3b ) 42246 DR311 (gRNA targeted to zebrafish ...gene rgs4 ) 42247 DR317 (gRNA targeted to zebrafish gene th1 ) 42248 DR330 (gRNA targeted to zebrafish gene... CRISPR Joung Lab CRISPR Plasmids Zebrafish gRNA CRISPR-Cas/RGN expression plasmids for Zebrafish...You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The Joung Lab recently...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...Publication Empty gRNA Expression Vectors You can use the tables on Addgene's Empty gRNA Vectors page to...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Chen pU3-gRNA OsU3 AarI none S. pyogenes Gao pZmU3-gRNA maize U3 none S. pyogenes Gao pU6-gRNA wheat U6...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...Selectable Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ....1016/j.bbrc.2013.08.013. Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...) at a specific location based on a co-expressed gRNA-defined target sequence. DSBs are preferentially...surrounding the DSB is introduced along with the Cas9 and gRNA plasmids, the cell may instead repair the break ...single category (cut, nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Type Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the...single category (cut, nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the...

... A gRNA to GFP 41820 gRNA_GFP-T2 A gRNA to GFP 41821 gRNA_DNMT3a-T1 A gRNA to DNMT3a 41822 gRNA_DNMT3a-T2...synthesizing gRNAs: hCRISPR gRNA synthesis protocol 115.2 KB A genome-wide resource of ~190k unique gRNAs targeting...41817 gRNA_AAVS1-T1 A gRNA to human AAVS1 41818 gRNA_AAVS1-T2 A gRNA to human AAVS1 41819 gRNA_GFP-T1 A...gRNA_DNMT3a-T2 A gRNA to DNMT3a 41823 gRNA_DNMT3b A gRNA to DNMT3b Yeast System: Table 2 To function in yeast cells...41824 gRNA Empty Vector The backbone that a target sequence gets cloned into to create the gRNA 41815 ...as a gRNA expression construct using the SNR52 snoRNA promoter. A protocol for synthesizing gRNAs: hCRISPR... details. A protocol for synthesizing gRNAs: Cas9 orthologs gRNA cloning protocol 104.1 KB This set of...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...genome. gRNA Empty Vectors Many gRNA empty vectors have been deposited at Addgene. To find the gRNA vector...Empty gRNA expression vectors for insertion of custom gRNA target sequences. CRISPR...target-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and a Cas ...pyogenes Puro Zhang phH1-gRNA 53186 Mammalian BbsI none S. pyogenes Gersbach pmU6-gRNA 53187 Mammalian BbsI...pyogenes Gersbach phU6-gRNA 53188 Mammalian BbsI none S. pyogenes Gersbach ph7SK-gRNA 53189 Mammalian BbsI...(U6a-BsaI-gRNA) 65955 Other/Tribolium castaneum BsaI none S. pyogenes Averof p(U6b-BsaI-gRNA) 65956 Other...

...Design homepage. gRNA plasmids ID Name Description PI 49408 pCFD1-dU6:1gRNA expression of gRNA under control...73914 pCFD5 multiplex gRNA plasmid Bullock 73915 pCFD6 tissue-specific multiplex gRNA plasmid Bullock Cas9... U6:1 promoter Bullock 49409 pCFD2-dU6:2gRNA expression of gRNA under control of the Drosophila U6:2 promoter... promoter Bullock 49410 pCFD3-dU6:3gRNA expression of gRNA under control of the Drosophila U6:3 promoter...promoter Bullock 49411 pCFD4-U6:1_U6:3tandemgRNAs expression of two gRNAs from Drosophila U6:1 and U6:3 Bullock...You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts Background CRISPR...Drosophila, consisting of transgenic cas9 fly lines and gRNA plasmids. Project tools can be used for both generating...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...multiple gRNAs that tile the locus. Chromosome painting requires delivery of hundreds of gRNAs with target...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Using catalytically...fluorescence microscopy in living cells. Alternatively, gRNAs can be fused to protein-interacting RNA aptamers...it is easy to implement due to the simplicity of gRNA design, programmable for different genomic loci,...efficiently visualized in living cells using a single gRNA that has multiple targeted sequences in close proximity...

...of two gRNAs in a single transcript whose expression is driven by a U6 promoter. The two gRNAs expressed...You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts This page lists plasmids...RFNs requires the expression of two guide RNAs (gRNAs) and a FokI-dCas9 fusion protein. The pSQT1313 plasmid...-specific Csy4 ribonuclease. Vectors expressing gRNAs targeted to sequences of interest can be made by...oligonucleotides to be synthesized for making specific gRNA pairs. ZiFiT Targeter is freely available online...the Csy4 ribonuclease (required for processing of gRNAs co-expressed in vectors made from pSQT1313) and ...links below: ID Plasmid 53370 pSQT1313 (multiplex gRNA expression plasmid) 53369 pSQT1601 (optimized Csy4...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...genomic DNA bound by the gRNA. An epitope tag(s) can be fused to dCas9 or gRNA for efficient purification...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Identifying molecules...of any genomic sequence specified by a particular gRNA. In the enChIP (engineered DNA-binding molecule-...vivo. For CRISPR-based purification, design your gRNA sequence to direct dCas9 to a specific locus, avoiding...

... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...CRISPR Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Catalytically dead...expression by interfering with transcription. Design your gRNA to target your gene of interest’s promoter/enhancer...

...interest 61079 gRNA-hIRF-1 #12 Expresses a guide RNA (gRNA) to target human IRF-1 promoter 61080 gRNA-hIRF-1 ...regions of interest. 98134 hIRF-1 gRNA #409 (Sa) Expresses a guide RNA (gRNA) to target human IRF-1 promoter...Sa-Cas9 system. 105283 hIRF-1 gRNA #351 (Sa) Expresses a guide RNA (gRNA) to target human IRF-1 promoter...interest 85586 gRNA Cloning Vector Bbs I ver. 2 An empty sgRNA expression vector. sgRNA sequence can be...CRISPR Cas9 gRNA plasmids from Dr. Hodaka Fujii's lab. Genome...You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts Originally developed...catalytically inactive form of Cas9 (dCas9) plus guide RNA (gRNA) for biochemical purification (Fig. 2). For additional...