...codon-optimized Cas9 constructs 44249 pdCas9-bacteria A catalytically inactive bacterial Cas9 expression plasmid...machineries, requiring only a nuclease-deficient Cas9 (dCas9) protein and a customized s ingle g uide RNA ...first plasmid ( pdCas9_bacteria ) contains an anhydrotetracycline (aTc)-inducible dcas9 gene on a p15A ...knockdown The first plasmid ( pdCas9_humanized ) contains a human codon optimized dcas9 gene under the control...Description 44246 pdCas9-humanized A catalytically inactive, human codon-optimized Cas9 expression plasmid...plasmid 44247 pdCas9::BFP-humanized A catalytically inactive, human codon-optimized Cas9-BFP fusion expression... plasmid 44250 pwtCas9-bacteria A wild-type Cas9 expression plasmid for use editing bacterial genomes ...

...chamber. Use the metal spatula to gently break the gel cast open. Use a razor blade to cut the top of the wells...

...consisting of a catalytically inactive form of Cas9 (dCas9) plus guide RNA (gRNA) for biochemical purification...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...regions of interest 51259 3xFLAG-dCas9/pMXs-I2 Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis...regions of interest 51260 3xFLAG-dCas9/pMXs-neo Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis...human IRF-1 promoter 62190 3xFLAG-dCas9/pTEF1p-CYC1t Expresses 3xFLAG-dCas9 in budding yeast for enChIP analysis...

...Root Cas9 Lentivirus Cas9 viruses can be used to interrogate the genome in a variety of ways. Cas9 proteins... 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...Resistance Activity PI 61422 dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A ...lenti dCAS-VP64_Blast dCAS9 (D10A, N863A) Blasticidin 3rd generation lenti vector encoding dCAS9-VP64 ...-targeting controls. This backbone contains SpCas9. SpCas9 and 76,441 unique sgRNAs targeting 19,114 human...plasmids resource page . Cas9 for Editing Genomic Sequences To perform genome edits, Cas9 nuclease is used to...

...Anti-CASPR/Neurexin IV [K65A/2R] CASPR/Neurexin IV Mouse IgG2a 206606 Anti-CASPR2 [K67/11R] CASPR2 Human...190491 CASK scFv [K56A/50] K56A/50 scFv CASK Human Mouse 190492 CASK scFv [K56A/57] K56A/57 scFv CASK Human...177445 Anti-CASPR/Neurexin IV [K65/35R] CASPR/Neurexin IV Rat Mouse IgG2a 177446 Anti-CASPR2 [K67/25R] ...219448 Anti-CASPR/Neurexin IV [K65A/30R] CASPR/Neurexin IV Rat Mouse IgG2a 219449 Anti-CASPR/Neurexin IV...SAP97 Rat Mouse IgG2a 128625 Anti-CASPR/Neurexin IV [K66/38R] CASPR/Neurexin IV Rat Mouse IgG2a 128626...Pan-KChIP K+ channel Mouse IgG2a 177443 Anti-CASK [K56A/50R] CASK Human Mouse IgG2a 177444 Anti-KChIP2b K+...Synaptotagmin-6 Mouse Mouse IgG2a 188217 Anti-CASK [K56A/57R] CASK Human Mouse IgG2a 188218 Anti-Kv2.1 K+ channel...

... for shuttling U6-guide cassettes to make dual guide expressing plasmids dCas9-KRAB-MeCP2 fusion for gene...enhancer sequence repression dCas9-VPR fusion for gene activation CRISPR/Cas9 Plasmid Variants Figure 1:...175574 express dCas9-KRAB-MeCP2 for repression. Plasmids 175571 and 175572 express dCas9-VPR for activation...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...plasmids to insert a second guide within a U6-guide cassette. This process is compatible with all pX458 family...Sources Original pX458 backbone ( Addgene #48138 ) dCas9 + KRAB-MeCP2 ( Addgene #110821 ) VPR ( Addgene #63798...

... Prepare one to two more conical tubes on ice in case you need to spread out the harvested cells further...

...links below: ID Plasmid 46294 pBS-Hsp70-Cas9 : A codon-optimized Cas9 nuclease under the control of the Drosophila...Drosophila hsp70 promoter. 45945 pHsp70-Cas9 : The codon-optimized Cas9 nuclease under the control of the Drosophila...Browse CRISPR Cas9 gRNA plasmids for drosophila research from the O'Connor-Giles, Wildonger, and [email protected] We have found that a variety of Cas9-mediated genome modifications can be efficiently...alterations can be generated within a month. phsp70-Cas9 and a single chiRNA plasmid can be coinjected to...imperfectly repaired DSBs. Coinjection of phsp70-Cas9 and plasmids encoding two chiRNAs can induce defined...engineering of Drosophila with the CRISPR RNA-guided Cas9 nuclease. Gratz SJ, Cummings AM, Nguyen JN, Hamm...

...contains Cas9. ID Plasmid Expression System Promoter Guide RNA Type Cloning Enzyme Co-expressed Cas9 Selection...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...transcriptase (RT) is fused to the C terminus of Cas9 H840A nickase. The fusion enzyme is capable of installing...template. During prime editing, the pegRNA directs the Cas9 nickase to the target sequence where it nicks the...nuclear localization signals, and two mutations in Cas9 to enhance nuclease activity; can be used in PE2...PE6 - small prime editors with optimized RT and/or Cas9 domains PE7 - addition of RNA-binding exonuclease...PuroTK-pegRNA_GG Mammalian, piggyBac hU6 pegRNA BsmBI Yes (Cas9 H840A + MMLV RT) PuroTK Jacob Giehm Mikkelsen 174038...

...inactive dCas9 is used to purify genomic DNA bound by the gRNA. An epitope tag(s) can be fused to dCas9 or ... for enChIP, as well as an anti-Cas9 antibody. Biotin tagging of dCas9 can be achieved by fusing a biotin...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...

...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors...require the use of lentivirus to deliver the gRNA/Cas9 to target cells. Therefore, you must be equipped...Doench and Root 2nd 2 40,710 Inzolia Human CRISPR/Cas12a Multiplex Knockout 209551 209552 Knockout Human... and Root 3rd 3–6 67,366 (Set A) 67,194 (Set B) Cas13a/C2c2 Protospacer flanking site (PFS) Library 79153...86550 Inhibition Human Weissman 3rd 10 Varies CRISPR/Cas9-assisted Removal of Mitochondrial DNA (CARM) Library... Sabatini and Lander 3rd 50 6,661 Garnett Lab MinLibCas9 Library 164896 Knockout Human Garnett 3rd 2 37,722...Human Stiewe N/A Varies 1,910–3,034 per exon Human Cas13 lncRNA Library 227014 RNA Knockdown Human Sanjana...

... protein dCas9 is expressed along a CRISPR array that contains 20 bp "addresses", where dCas9 is going...going to bind on the chromosome. When dCas9 binds to a gene, it blocks the progression of the RNA polymerase...for quick integration of the aTc-inducible pTet-dCas9 in the attachment site of the phage 186. Detailed... Bikard, D., & Van Teeffelen, S. (2018). Tuning dCas9’s ability to block transcription enables robust,...Schematic of the strain expressing two reporters and dCas9 under a P-tet promoter integrated in the chromosome... of the two reporters, mCherry and sfgfp in this case, can be controlled using a plasmid‐borne CRISPR ...

...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...

...familial ALS cases having at least one other affected family member. These familial cases, usually inherited...range of cellular processes. The vast majority of cases (90-95%) of ALS are sporadic, having no prior family...cause) nature of the disease, a small portion of cases can be attributed to known genetic factors such ...early-onset form accounts for less than 10 percent of all cases of Alzheimer's disease. Of these early-onset forms...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...

...domain death agonist CASP3 Caspase 3, apoptosis-related cysteine peptidase CASP8 Caspase 8, apoptosis-related...apoptosis-related cysteine peptidase CASP9 Caspase 9, apoptosis-related cysteine peptidase Cyclin B CCNB1 CCNB2 CCNB3...

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

...interest should be active, and only turned off occasionally, using tetracycline off or tTA is more appropriate...should be mostly inactive, and only turned on occasionally, using tetracycline on or rtTA is more appropriate...inducible expression for Gateway cloning; selection cassette in format: PGK-rtTA-2A-puro; see article for tagged...inducible expression of a miR-shRNA; selection cassette in format: rtTA3+IRES+Neo; see article for additional...conditional cell line generation; contains expression cassette for TetR-P2A-GFP; see article for additional constructs... rtTA On Alexeyev 14901 pCL-CTIG The inducible cassette contains tTA at its 5' end and an inducible promoter...

...mE10 CARD11 caspase recruitment domain family, member 11 BIMP3, CARMA1, MGC133069 CBL Cas-Br-M (murine...retroviral transforming sequence C-CBL, CBL2, RNF55 CBLB Cas-Br-M (murine) ecotropic retroviral transforming sequence...DKFZp779F1443, FLJ36865, FLJ41152, Nbla00127, RNF56 CBLC Cas-Br-M (murine) ecotropic retroviral transforming sequence...BRAF1, FLJ95109, MGC126806, MGC138284, RAFB1 CASP3 caspase 3, apoptosis-related cysteine peptidase CPP32...ASBT, ISBT, NTCP2 TAP1 transporter 1, ATP-binding cassette, sub-family B (MDR/TAP) ABC17, ABCB2, APT1, D6S114E...TAP1*0102N, TAP1N TAP2 transporter 2, ATP-binding cassette, sub-family B (MDR/TAP) ABC18, ABCB3, APT2, D6S217E...IMD1, MGC126261, MGC126262, PSCTK1, XLA CARD11 caspase recruitment domain family, member 11 BIMP3, CARMA1...

...typically arise spontaneously, although in rare cases Rett syndrome can be inherited. In females, random...R306C) account for almost 46% of all Rett syndrome cases. Research Milestones 1992 - MECP2 is discovered ...Knock-in of NLuc-tdTomato at endogenous MECP2 locus Castaneus MECP2-NLuc-tdTomato mouse reporter cell line (...resources for a full selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples of additional...hand use in girls: Rett’s syndrome: report of 35 cases. Ann Neurol . 14, 471–479. (Link opens in a new ...

...markers. The FP together with the protothropic cassette can be amplified with the following primers: FW...Yeast Localization and Overexpression (MyLO) CRISPR-Cas9 Toolkit Other Plasmids Bas Teusink Lab Materials...in a new window) Sheff MA, Thorn KS. Optimized cassettes for fluorescent protein tagging in Saccharomyces...Bydder SF, Zhou Y, Nielsen LK. Dual gene expression cassette vectors with antibiotic selection markers for ...