...PubMed . Return to AAV In vivo plasmids AAV Plasmids with Cas9 Mouse CRISPR-Cas9 knockin mice for genome editing...pAAV-pMecp2-SpCas9-spA) for expressing Cas9 in neurons. SpCas9 guide acceptor: The PX552 plasmid (#60958)...tracrRNA: This plasmid contains three expression cassettes. To target a given site, the plasmid can be digested...Return to top Cas9 SAM transcriptional activation plasmids and screening library CRISPR/Cas9 Synergistic...constitutively expressing Cas9 knockin mice (Platt et al ., Cell 2014). In these mice the CRISPR-Cas9 system ...constitutively Cas9-expressing mouse. Described here are AAV vectors that can be combined with Cas9 in a wide...information: CRISPR-Cas9 mouse toolbox protocol 637.8 KB CRISPR-Cas9 Cre expression vectors for cancer ...

...and/or Cas9 expression or (2) a lack of efficient target cleavage in cells expressing both Cas9 and gRNA...the multi-plasmid SAM system. Select Expression System To use CRISPR, you will need both Cas9 and a gRNA...for details, see Plasmids 101: Mammalian Vectors ) Transient or stable expression of Cas9 and/or gRNA in...organism without a specific preferred mutation Cas9 (or Cas9 nickase) Single (or dual) gRNA targeting 5′ ...generating a point mutation or inserting a tag Cas9 (or Cas9 nickase); Base editor Single (or dual) gRNA ...concern, Cas9-gRNA ribonucleoprotein (RNP) complexes are advantageous due to the transient Cas9 expression...Delivery method Expression System Components of System Application Mammalian expression vector Cas enzyme...

...Whether the plasmid contains Cas9 and if so, which function of Cas9. We have plasmids with Cas9 that can ...by the Doudna Lab . Plasmid Expression System Cloning Enzyme Co-expressed Cas9 Cas9 System Selection PI... on the plasmid. gRNA plasmids that do not co-express a Cas protein require a separate plasmid that does...be used to express multiple gRNAs from a single plasmid with or without expression of Cas9. Highlighted.... Can express Cas9 with up to 3 gRNAs. Frew pSQT1313 Mammalian Single plasmid for the expression of multipe...Mammalian Lentiviral expression of Cas9, dCas9, or dCas9-VP64 along with 1-4 sgRNAs expressed from independent...Vectors CRISPR Plasmids: Empty gRNA Expression Vectors Browse CRISPR Plasmids By Function Genome Editing Cut...

...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...none, need Cas9 plasmid Boxem 47941 pMB60 T7 BsaI In vitro transcription none, need Cas9 plasmid Boxem 47912...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...need Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid... elegans none, need Cas9 plasmid Ristow Do you have suggestions for other plasmids that should be added...specific locus. ID Plasmid Gene/Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA... gRNA expression plasmid based on factors such as selectable marker or cloning method. When using CRISPR...

...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...transcription Virmilion none, need Cas9 plasmid Bullock and Port 49330 pAc-sgRNA-Cas9 dU6 BspQI Transfection Puromycin..., need Cas9 plasmid Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid...CRISPR Plasmids: Drosophila Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime...homology-directed repair (HDR). ID Plasmid Gene/Insert Promoter PI Publication Prime Edit Cas9 H840A nickase fused to...specific locus. ID Plasmid Gene/Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA...

... Plasmid Collections Worm Plasmids Worm Plasmids and Resources Plasmid Tools Worm Constructs...some of the plasmids and tools available at Addgene for use in worms. Browse Plasmids Plasmid Tools for ... Gene/Insert PI Plasmids Expressing Worm Genes/Inserts The table below lists plasmids that contain a worm...Addgene's worm CRISPR resources include plasmids expressing Cas9, empty gRNA backbones, and constructs ... genes by feeding worms bacteria expressing dsRNA or use CRISPR/Cas9 gene editing to easily engineer specific...backbones, fluorescent proteins, and Cre expression plasmids. Large collections or kits are described ...complete list of plasmids that can be used in worms, search Addgene's entire collection. ID Plasmid Description...

...high expression, so adding this G could help with expression, though it is possible for the plasmid to ...double nickase, you could express two spacers and use PX335 to express the Cas9n (nickase). The concept ...need to have four spacers co-expressed and using PX335 to express the Cas9n (nickase). Homologous Recombination... may also like... Zhang Lab CRISPR plasmids General CRISPR Plasmids Dr. Le Cong, the first author of one...nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase which will together...the Cas9 nickase doesn't have the ability to induce double-stranded breaks like the wildtype Cas9 does...easiest way is to co-transfect several plasmid together, with each plasmid having a targeting spacer cloned ...

...codon-optimized Cas9-BFP fusion expression plasmid 44248 pgRNA-humanized A customizable gRNA expression plasmid... inactive, human codon-optimized Cas9 expression plasmid 44247 pdCas9::BFP-humanized A catalytically inactive...codon-optimized Cas9 constructs 44249 pdCas9-bacteria A catalytically inactive bacterial Cas9 expression plasmid...plasmid 44250 pwtCas9-bacteria A wild-type Cas9 expression plasmid for use editing bacterial genomes 44251...customizable gRNA expression plasmid for use with the bacterial Cas9 constructs 46910 pHR-SFFV-dCas9-BFP Human...1. Two-plasmid CRISPRi system for bacterial gene knockdown The first plasmid ( pdCas9_bacteria ) contains...2. Two-plasmid CRISPRi system for mammalian gene knockdown The first plasmid ( pdCas9_humanized ) contains...

... Plasmid Collections Plant Plasmids Plant Plasmids and Resources...for a full selection of plasmids expressing different functional variants of Cas9 and empty gRNA backbones...creating plant expression vectors: A modular plasmid assembly kit for multigene expression, gene silencing...addition to individual plasmids, Addgene carries several useful toolkits and plasmid collections for use ...includes plasmids containing promoters that drive general and species-specific gene expression (see the... assembly of one or more gRNA expression cassettes - Qi-Jun Chen. Cas9 and gRNA backbone for use in rice...CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants - Niko Geldner. CRISPR-Cas9 vector...

... 52962 lentiCas9-Blast Sp Cas9 Cut Blasticidin Expresses human codon-optimized S. pyogenes Cas9 protein...backbone. Zhang Cas9 and Accessories for Activating Gene Expression Catalytically-dead Cas9 (dCas9) can be fused...Root Cas9 Lentivirus Cas9 viruses can be used to interrogate the genome in a variety of ways. Cas9 proteins...CRISPR plasmids resource page . Cas9 for Editing Genomic Sequences To perform genome edits, Cas9 nuclease...Libraries Cas9 Pooled Barcoding Libraries Control Addgene's lentiviruses are made from plasmids that are... list of gRNA plasmids targeting specific genes. For more information on CRISPR plasmids and function,... of available viruses based on plasmids in our repository that express fluorescent proteins. ID Name Insert...

...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...transcription none, need Cas9 plasmid Chen Do you have suggestions for other plasmids that should be added...following CRISPR plasmids have been designed for use in Xenopus. Cut Fully functional Cas9 enzymes designed...surrounding the DSB is introduced along with the Cas9 and gRNA plasmids, the cell may instead repair the break ...genomic changes. ID Plasmid Gene/Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA... gRNA expression plasmid based on factors such as selectable marker or cloning method. When using CRISPR...nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with a ...

...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In Resistance Co-expressed Cas9 Depositing...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert....2013.08.013. Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors such as ...Ampicillin none, need Cas9 plasmid Qi Do you have suggestions for other plasmids that should be added ...population. ID Plasmid Promoter PI Publication Purify A catalytically inactive Cas9 (dCas9) can be used ...Kanamycin none, need Cas9 plasmid Marraffini 42876 pCas9 BsaI E. coli, S. pneumoniae Chloramphenicol yes, cut...

...CRISPR-Cas/RGN expression plasmids for use in human cells CRISPR-Cas/RGN expression plasmids for use in zebrafish...zebrafish Guide RNA expression plasmids for the EGFP Reporter Gene Guide RNA expression plasmids for endogenous...reagents: Cpf1 expression plasmids (AsCpf1 and LbCpf1) for use in mammalian cells SpCas9 variants with ...specifity of Staphylococcus aureus Cas9 (SaCas9 KKH variant) Engineered CRISPR-Cas9 nucleases with altered PAM...the Cas9 endonuclease) that cleaves the target DNA. The Joung lab recently described gRNA and Cas9 expression...endogenous human genes Guide RNA expression plasmids for endogenous zebrafish genes...RNA-Guided Nuclease (RGN) Plasmids Genome...

...Browse Plasmids: Purify Cas9 Alternatives for CRISPR Genome Engineering S. pyogenes Cas9 (SpCas9) is the...more about Cas9 multiplexing . Browse Plasmids: Multiplex gRNA Vectors Engineering the Cas9 Endonuclease... available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...in which Cas9 is included in the gRNA-containing plasmid, or 2-vector systems, in which Cas9 must be delivered...to either co-infect with a Cas9-producing lentivirus or use a Cas9-expressing stable cell line. After applying...strand SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 - increase Cas9 proofreading...new tools. Figure 3: Overview of Cas9n Nickase and dead Cas9 The two Cas9 nuclease domains, RuvC and HNH...

...vectors using ssDNA oligos p201G Cas9 ; p201B Cas9 ; p201H Cas9 ; p201N Cas9 ; pUC gRNA Shuttle PDF 488.6 ...CRISPR RNA array: Cas9 (pX260) or Cas9 D10A (pX334) ; tracrRNA: Cas9 (pX330) or Cas9 D10A (pX335) ; PDF...KB Church gRNA design and cloning for Cas9 orthologs Cas9 plasmids PDF 104.1 KB Chen and Wente Zebrafish... CRISPR plasmid summary for transcriptional activation dCas9 activators sgRNA dual expression: pAC2 , ...design and cloning All-in-one plasmid containing expression cassette for NmCas9 and BsmBI site for cloning...putative target sites for S. pyogenes Cas9, S. thermophilus Cas9, or Cpf from your input sequence or list...vitro transcription, injection gRNA core ; Cas9 ; optimized Cas9 PDF 66.8 KB Fujii gRNA design and cloning...

... Plasmid Collections Zebrafish Plasmids Zebrafish Plasmids and Resources... of the plasmids and tools available at Addgene for use in zebrafish. Browse Plasmids Plasmid Tools for...biosensors, and Cre expression plasmids. Large collections or kits are described below . ID Plasmid Description...CRISPR resources for a full selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples...Using CRISPR/Cas9 - Jeffrey Essner Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) expression plasmids - Keith Joung...for plasmids containing your zebrafish gene of interest? Search Addgene's collection for plasmids that...highlights plasmids that contain tools for use in zebrafish research. This includes plasmids for genome...

...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing...need Cas9 plasmid Chen and Wente 42250 DR274 T7 BsaI In vitro transcription none, need Cas9 plasmid Joung...than NHEJ. ID Plasmid Gene/Insert Vector Type Promoter PI Publication Empty gRNA Expression Vectors Select...Select a gRNA expression plasmid based on factors such as selectable marker or cloning method. When using...nick, etc.) On the other hand, gRNA plasmids that do not co-express a Cas protein can be paired with a ...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...CRISPR plasmids that have been designed for use in zebrafish. CRISPR...

...Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In Resistance Co-expressed Cas9 Depositing...also express a gRNA, you will need to use a separate gRNA expression plasmid to target the dCas9-activator...separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert...cerevisiae LEU2 none, need Cas9 plasmid Lu Do you have suggestions for other plasmids that should be added ...of the coding sequence. If the plasmid you’re using does not also express a gRNA, you will need to use ...sites. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Empty gRNA Expression Vectors Select... Select a gRNA expression plasmid based on factors such as selectable marker or cloning method. When ...

...selectable marker, and whether the plasmid contains Cas9. ID Plasmid Expression System Promoter Guide RNA Type...below for CRISPR prime editing plasmids. Plasmids are available for expression in mammalian systems, bacteria...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Drosophila ID Plasmid Gene/Insert...Type Cloning Enzyme Co-expressed Cas9 Selection PI 132777 pU6-pegRNA-GG-acceptor Mammalian hU6 pegRNA ...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...prime editor plasmids for (search and replace) genome editing. CRISPR...

...below for CRISPR base editing plasmids. Plasmids are available for expression in mammalian systems, bacteria...are created by fusing Cas9 nickase or catalytically inactive “dead” Cas9 (dCas9) to a cytidine deaminase...CRISPR Plasmids: Base Edit Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime Edit...editors are available with multiple Cas9 variants including high fidelity Cas9’s. Further advancements have ...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Yeast ID Plasmid Gene/Insert...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...