...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...

...Nathaniel Roquet from the Lu Lab at MIT developed plasmid based systems that report the order in which specific... action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (AAVs) are... editing - all influenced by an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking “... plasmid... system that uses modular RNA binding proteins, dCas9, and a modified gRNA to direct a variety of customizable...

...interest (or the whole genome), they knock down gene expression temporarily. RNAi libraries are especially useful... Genetic screens in human cells using the CRISPR/Cas9 system. Science, 343(6166): 80-84 (2014). 10.1126...

...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...

...   It’s not an overstatement to say that CRISPR/Cas9 editing is one of the key tools now providing access...plans and in particular, that they all share the plasmids created under this programs via Addgene. New organisms...snail can be manipulated by injected RNA and can express fluorescent proteins. This lab has created an automated...sexual reproductive life cycles. One can inject plasmid DNA in the eggs to get transgenic animals. They...-Mediated Ovary Transduction of Cargo)  delivers Cas9 RNP to the arthropod germline by injection into ...

...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...

...has a low quantum yield, and when transiently expressed in cells, generates much less signal than comparable... endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent signal for imaging ...: 1262. PMID: 23232392 PMCID: PMC3535334 Find plasmids from this paper at Addgene. Takai, Akira, et ...4352-6. PMID: 25831507. PMCID: PMC4394297 Find plasmids from this paper at Addgene. Suzuki, Kazushi, ... 13718. PMID: 27966527 PMCID: PMC5171807 Find plasmids from this paper at Addgene. Additional Resources...All Our Fluorescent Protein Resources Find FRET Plasmids Find Empty Backbones with Fluorescent Protein ...

...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...

...Browse fluorescent protein plasmids and resources See all Jaffrey lab plasmids ... cellular processes by transforming cells with plasmids encoding aptamer-tagged RNA and later introducing...aptamer downstream of a known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the enrichment...leading to conformational changes that control the expression of coding RNA. Spinach-based riboswitches can...cellular RNA to specific intracellular locations using Cas9 and track the localization of the complex. Broccoli...SELEX, the RNA pool was cloned into bacterial expression vectors to create a library. Bacteria were transformed...quantify RNA transcription by RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely...

...generation of multiple edits or implementation of expression systems to accurately mimic a disease phenotype...CRISPR 101: Making a Knock-In Cell Line Using CRISPR-Cas9 to Edit Disease Out of the genome Tips for a first...resources on Addgene.org CRISPR Guide CRISPR Plasmids and Resources CRISPR History and Development ...

...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...

...Addgene plasmid ID 42230). This plasmid allows for the simultaneous expression of sgRNA and SpCas9, but ...cloning and expression purposes. NOTE: Here we discuss the usage of the pSpCas9(BB) plasmid (pX330) (Addgene...utilized, such as pX458 (Addgene plasmid ID 48138) or pX459 (Addgene plasmid ID 48139), which include GFP ...constructs in which multiple sgRNAs may be expressed from a single plasmid. Add “CACC” before the 20-mer guide...This protocol involves the delivery of CRISPR/Cas9 plasmids by electroporation 4 . This protocol is described.... However, transient transfection of CRISPR/Cas9 plasmids may be successfully adapted to numerous cell...Add 5 μg of each CRISPR/Cas9 construct (10 μg total). Add 0.5 μg of GFP expression construct. Electroporate...

...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...CRISPR Plasmids: Mammalian Expression The following CRISPR plasmids have been designed for use in mammalian...FLAG-tagged Cas9. In the CAPTURE system, Cas9 is tagged with a biotin acceptor site and co-expressed with BirA...NHEJ. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Browse CRISPR Plasmids By Function...repair (HDR). ID Plasmid Gene/Insert Promoter Selectable Marker Publication Prime Edit Cas9 H840A nickase...of the coding sequence. If the plasmid you’re using does not also express a gRNA, you will need to use ...

...Tagging Plasmids: pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA...and Myers Tagging Plasmids Protein Species Tag Donor Plasmid gRNA plasmid gRNA plasmid FLAG pFETCh_Donor...the remaining 5 plasmids include Cre and FLP expression vectors, a general cloning plasmid, and a prebuilt...Doyon lab, eSpCas9(1.1)_No_FLAG_AAVS1_T2 (Addgene #79888) , which expresses an untagged Cas9 AND a gRNA... This CRISPR/Cas plasmid-system consists of two components: A vector containing Cas9 and a validated gRNA.... Multiple gRNA plasmids may be used. The HDR (homology directed repair) donor plasmid with homology arms...well as pX330S-2-PITCh, expressing a generic gRNA targeting the PITCh donor plasmid. Note that the principle...

...different plasmids. Plasmids within a given library have the same backbone, but they express or target... Science Guides Guide to Plasmid Pooled Libraries Plasmid Pooled Libraries Jump To: Pooled Library...libraries, each plasmid contains a unique cDNA. In shRNA or gRNA libraries, each plasmid contains a unique... others, notably pooled lentiviral plasmid libraries, the plasmids must first be used to make virus. This...infected cell receives only one plasmid. To make sure that every plasmid is adequately represented in the... given plasmid, think of each cell carrying it as being a biological replicate for that plasmid. Library...the overall library. Barcoding libraries contain plasmids with unique, semi-random sequences that can be...

...Article Individual plasmids can be ordered via the links below: ID Plasmid 46294 pBS-Hsp70-Cas9 : A codon-optimized.... (2013). Plasmid is low copy number. 45946 pU6-BbsI-chiRNA : Plasmid for expression of chiRNA under the...Browse CRISPR Cas9 gRNA plasmids for drosophila research from the O'Connor-Giles, Wildonger, and Harrison... CRISPR flyCRISPR Lab CRISPR Plasmids flyCRISPR Plasmids Available from Addgene You may also like...be generated within a month. phsp70-Cas9 and a single chiRNA plasmid can be coinjected to generate mutations...imperfectly repaired DSBs. Coinjection of phsp70-Cas9 and plasmids encoding two chiRNAs can induce defined deletions...These plasmids are described in: Genome engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease...

...Turquoise (CeNL; Plasmid #85199), Yellow (YeNL; Plasmid #85201), Orange (OeNL; plasmid #85202) and Red ... Researchers have been sharing plasmids ever since there were plasmids to share. Back when I was in the...specific strategies and tactics for getting the plasmids he wanted. Addgene makes it so much easier to ...endogenous tagged genes generated through CRISPR/Cas9 knock-in. Finally, NLuc is very inviting for endogenous...kDa. An example is the CRISPaint-NLuc construct (Plasmid #67178) for use in the system outlined in Schmid-Burgk...reporter was used to sort successfully infected/expressing cells by flow cytometry. The isolated cells were...CyOFP1-NLuc-CyOFP1; Antares). Direct injection of plasmids for Antares or firefly luciferase into mouse livers...

...and 175574 express dCas9-KRAB-MeCP2 for repression. Plasmids 175571 and 175572 express dCas9-VPR for activation...shuttle plasmid for shuttling U6-guide cassettes to make dual guide expressing plasmids dCas9-KRAB-MeCP2...guides can be expressed from the same plasmid by utilizing one of the dual guide shuttle plasmids to insert...enhancer sequence repression dCas9-VPR fusion for gene activation CRISPR/Cas9 Plasmid Variants Figure 1: Schematic...advantage of the Zhang lab all-in-one (guide/Cas9/reporter) pX458 plasmid family. They have made several modifications... the CRISPR Collection. Plasmids 159654 and 159655 are single guide expressing double stranded break constructs...constructs. Plasmids 171098 and 175570 allow conversion to a dual guide construct. Plasmids 175573 and ...

...dystrophy. While mini-dystrophin expression levels were lower than expression of wild-type dystrophin in normal...Drawbacks: Transgene expression varies with this approach, ranging from ~10% expression in primary human ...this AAV successfully mediated transduction and expression of the full-length transgene in vitro and in ...genomes complement each other to restore the full-expression cassette. AAV’s natural ability to reassemble...overlapping split vector has equal or higher transgene expression when compared to using a single over-sized vector...could have a big impact on the rate of transgene expression with an overlapping AAV since both of these variables...sequences, the group achieved ~80% of muscle cells expressing mini-dystrophin.  3. Hybrid  Overview: The hybrid...