...Lab CRISPR/Cas9-based conditional mutagenesis in zebrafish - Wenbiao Chen Lab A CRISPR/Cas9 Vector System...modified CRISPR-Cas9 system - Shuo Lin Lab Re-purposed Ac/Ds transgenic system enabling CRISPR/dCas9-interference...morpholinos, zinc finger nucleases, TALENS, and CRISPR/Cas9-based gene editing make the zebrafish a stalwart...resources for a full selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples of useful...GeneWeld Vectors for Targeted Integration Using CRISPR/Cas9 - Jeffrey Essner Lab CRISPR-Cas/RNA-Guided Nuclease...use of zebrafish. CRISPRz - A database of CRISPR/Cas9 target sequences that have been experimentally validated...

...Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes (PAM = NGG) 46759...none, need Cas9 plasmid Chen and Wente 42250 DR274 T7 BsaI In vitro transcription none, need Cas9 plasmid...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

...) Validated In Resistance Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes (PAM = NGG) 49014... Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA...immunoprecipitate FLAG-tagged Cas9. Design your gRNA sequence to direct dCas9 to a specific locus, avoiding...handle_RPR1t pRPR1 HindIII S. cerevisiae LEU2 none, need Cas9 plasmid Lu Do you have suggestions for other plasmids...PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...Marker PI Publication Base Edit Catalytically dead dCas9 fused to a cytidine deaminase protein becomes a ...

...pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA, & FBL-specific...Doyon lab, eSpCas9(1.1)_No_FLAG_AAVS1_T2 (Addgene #79888) , which expresses an untagged Cas9 AND a gRNA... consists of two components: A vector containing Cas9 and a validated gRNA, based on the Zhang lab's PX458...need to design and clone in a gRNA(s) to guide the Cas9 protein to your target sequence, as well as design...consists of two components: An all-in-one CRISPR/Cas9 vector based on the Zhang lab's PX330 , simultaneously... to generate both an expression cassette for the Cas9-programming sgRNA and HR donors for selectable genome...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

...Tools CRISPR Blog Posts A catalytically inactive Cas9 (dCas9) is fused to FokI nuclease. When FokI dimerizes... CRISPR dCas9-FokI CRISPR: dCas9-FokI...with dCas9 fused to FokI for creating double-strand breaks (DSBs). CRISPR...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...gRNAs, binding ~15-25 bp apart, are required for dCas9-FokI to dimerize at a given region of the genome...

... to immunoprecipitate FLAG-tagged Cas9. In the CAPTURE system, Cas9 is tagged with a biotin acceptor site... Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of genomic DNA...Publication Visualize A catalytically inactive Cas9 (dCas9) fused to a fluorescent protein (FP) can help...Promoter Selectable Marker Publication Prime Edit Cas9 H840A nickase fused to a reverse transcriptase (...PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors... CRISPR Blog Posts Base Edit Catalytically dead dCas9 fused to a cytidine deaminase protein becomes a ...

...in retinal development the lab utilizes a CRISPR/Cas9 based approach to knockout, repress, and activate...taken advantage of the Zhang lab all-in-one (guide/Cas9/reporter) pX458 plasmid family. They have made several...fusion for gene activation CRISPR/Cas9 Plasmid Variants Figure 1: Schematic of the CRISPR Collection. Plasmids... plasmids dCas9-KRAB-MeCP2 fusion for gene and non-coding enhancer sequence repression dCas9-VPR fusion...175574 express dCas9-KRAB-MeCP2 for repression. Plasmids 175571 and 175572 express dCas9-VPR for activation...Sources Original pX458 backbone ( Addgene #48138 ) dCas9 + KRAB-MeCP2 ( Addgene #110821 ) VPR ( Addgene #63798...

... Plasmid Promoter Cloning Enzyme Co-expressed Cas9 Cas9 System Selection PI 46968 pICSL01009::AtU6p aU6...Promoter Selectable Marker PI Publication Prime Edit Cas9 H840A nickase fused to a reverse transcriptase (...BsaI yes, cut S. pyogenes Hygro Chen 78932 pHDE-35S-Cas9-mCherry-UBQ U6-26 PmeI yes, cut S. pyogenes Hygro...PI Publication Interfere Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...Marker PI Publication Base Edit Catalytically dead dCas9 fused to a cytidine deaminase protein becomes a ...Marker PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can...

...consisting of a catalytically inactive form of Cas9 (dCas9) plus guide RNA (gRNA) for biochemical purification...CRISPR Cas9 gRNA plasmids from Dr. Hodaka Fujii's lab. Genome...(gRNA) to target human IRF-1 promoter for the Sa-Cas9 system. 105283 hIRF-1 gRNA #351 (Sa) Expresses a...(gRNA) to target human IRF-1 promoter for the Sa-Cas9 system. References An enChIP system for the analysis...Description 47948 3xFLAG-dCas9/pCMV-7.1 enChIP plasmid, expresses 3xFLAG-dCas9 in mammalian cells for enChIP...in mammalian cells 51240 3xFLAG-dCas9/pMXs-puro Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis...regions of interest 51258 3xFLAG-dCas9/pMXs-IG Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis...

...species or PAM binding variant of Cas9. For instance, wild-type SpCas9 must be used with targets that are...increases off-target effect. Which species or variant of Cas9 ( S. pyogenes , S. aureus etc.) was this gRNA sequence...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

...genomic deletions. Compared to deletions generated by Cas9, large deletions created using Cas3 have fewer off-target...underutilized compared to their Class 2 counterparts like Cas9. This is attributed in part to the difficulties ...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

...single species or PAM binding variant of Cas9 (e.g. Wild-type SpCas9 must be used with targets that are upstream...activity. Know your target. Which species or variant of Cas9 ( S. pyogenes, S. aureus etc.) was this gRNA sequence...and Resources page have been used to indicate the Cas9 application the gRNA was designed to accomplish....Target Species Target Sequence Plasmid ID Application Cas9 Species PubMed ID Depositor OCT4 H. sapiens CTCCCATGCATTCAAACTG...10 C. elegans CGCTACCATAGGCACCACG 71516 cut VRER Cas9 mutant 26680661 Fire dpy-10 C.elegans GCTACCATAGGCACCACGAG...-10 C. elegans TCCGCTACCATAGGCACCA 71479 cut VQR Cas9 variant 26680661 Fire dpy‐10 C. elegans GCTACCATAGGCACCACGAG...Christiaen EGFP A. victoria multiple, see article 60071 dCas9-FokI S. pyogenes 24770325 Joung EGFP A. victoria...

...Cre-dependent Cas9: Cre-dependent Cas9 can be knocked in to a mouse at the Rosa26 locus to facilitate Cas9 mediated...iRFP670, Cre, and GFP EFS Lentiviral Jacks 68468 Cas9-2A-Cre Cas9 and Cre CAG Mammalian Zhang 68477 pTRE:iRFP670...sgRNA expression hSyn AAV Zhang 60820 pSECC Cre, Cas9 and sgRNA expression EF-1 alpha Lentiviral Jacks...

... for enChIP, as well as an anti-Cas9 antibody. Biotin tagging of dCas9 can be achieved by fusing a biotin...inactive dCas9 is used to purify genomic DNA bound by the gRNA. An epitope tag(s) can be fused to dCas9 or ...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...biotin acceptor site to dCas9 and co-expressing BirA biotin ligase, as seen in the CAPTURE system. The ...purification, design your gRNA sequence to direct dCas9 to a specific locus, avoiding known transcription...

...crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which require that each DNA target have...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

... inactive Cas9 (dCas9) fused to a fluorescent marker like GFP, researchers have turned dCas9 into a customizable...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...

...research for decades. Recent developments with CRISPR-Cas9 gene editing have propelled both reverse and forward...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...forward genetics of these parasites. Moreover, CRISPR-dCas9 gene regulation has also been established for some...

...Target neural oxytocin receptors using an AAV-CRISPR/Cas9 strategy for gene editing across divergent rodent...species. Boender et al. Sci Adv. 2023 Jun 2. Use Cas9 in astrocytes. Endo et al Science. 2022 Nov 4. Base...

...Technique PI Purpose pCas9 42876 CRISPR Luciano Marraffini Bacterial expression of Cas9 nuclease gRNA. For...Anhydrotetracycline inducible expression of wild-type Cas9 from S. pyogenes for inducing double stranded breaks. For...Anhydrotetracycline inducible expression of catalytically inactive Cas9 ( S. pyogenes ) which can be combined with expression...list of our bacterial CRISPR plasmids, see here . pwtCas9-bacteria 44250 CRISPR Stanley Qi Anhydrotetracycline...pPro24-gfp contains GFP but all others are empty. pdCas9-bacteria 44249 pTetO Anhydrotetracycline Stanley...

...Yeast Localization and Overexpression (MyLO) CRISPR-Cas9 Toolkit Other Plasmids Bas Teusink Lab Materials...