...genes using dCas9-based activators The simplest of the dCas9 activators consists of dCas9 fused to a single... Fig. 3: Cas9 repressors include dCas9 alone and a dCas9-KRAB fusion, both targeted to the promoter...Repressing target genes using dCas9-based repressors Early experiments using dCas9 in bacteria demonstrated...is frequently fused to Cas9 for this purpose. dCas9-KRAB plasmids for repressing target genes in a variety...even making a break.  Base editors are dead Cas9 (dCas9) or nCas9 enzymes fused to either cytosine or adenosine...genome but still want to affect gene expression? dCas9 can be used as a platform to deliver various cargo...specific DNA locus by directly fusing the cargo to dCas9.  This technique has been exploited to localize ...

...an EGFP reporter. Expression of the dCas9-KRAB activator or dCas9-VPR repressor in hPSCs is also driven...piggyBac gRNA delivery and the all-in-one dCas9-KRAB and dCas9-VPR targeting plasmids can be found at Addgene...of single or multiplexed gRNAs and of dCas9 fusions. The dCas-KRAB activator and the dCas-VPR repressor...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly... could quantify and track cells expressing both dCas9 and gRNAs. The multi-gRNA piggyBac vector ...plasmids for piggyBac gRNA delivery and the all-in-one dCas9 targeting plasmids! Hazelbaker et al., Sci Reports...

...including pcDNA-dCas9-p300 Core and pcDNA3.3-Nm-dCas9-p300 Core, as well as pLV-dCas9-p300-P2A-PuroR for...pCMV-dCas9-D3A) and Tet-dependent (TetO-dCas9-D3A) constructs. For lentiviral expression, Fuw-dCas9-Dnmt3a...Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase...Fusing dCas9 to a transcriptional activator like VP64 or VPR activates transcription, whereas dCas9-KRAB...activators/repressors - dCas9-p300 increased transcriptional activation more than dCas9-VP64, especially when...Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety of epigenetic modifiers to specific...Transcriptional activation p300 acetyltransferase dCas9 fused to the catalytic domain of p300 acetyltransferase...

... nickase, but we are now adopting dCas9, dCas9-VP64, and dCas9-KRAB in our system,” Sakuma said. “These...

...fluorescent dCas9-KRAB and dCas9-VPR for quantifying and tracking cells that express both the dCas9 construct...Saccharomyces cerevisiae. The system consists of dCas9, the mammalian transcriptional repressor Mxi1, and...accessible and the complex enters the nucleus where dCas9-Mxi1 can repress gene expression. In the dark, the...Caulobacter cells for ctrA gene expression using various dCas9. Image adapted from Guzzo et al., 2020. Caulobacter...Caulobacter genes using a catalytically dead Cas9 (dCas9) derived from Streptococcus pasteurianus  or the...essential cell cycle regulator. The lab expressed both dCas9 and the ctrA sgRNA and saw that the levels of ctrA...minutes in xylose (which controls expression of dCas9). The lab demonstrated the system can also be used...

...binding protein (dCas9). dCas9 is then tethered to a repressive domain, such as KRAB, to prevent efficient...loss of function while nuclease-deactivated Cas9 (dCas9) can be tethered to a repressor (CRISPRi) or an ...library In addition to inhibiting gene expression, dCas9 can also be used for transcriptional activation....developed to recruit transcriptional machinery to dCas9 and its RNA directed binding site. These methods... fusion of an activation domain such as VP16 to dCas9 or the recruitment of multiple copies of VP16 termed... with only two constructs (the other containing dCas9-VP16). Like CRISPRi, CRISPRa is only effective when...

...repression of protein expression. Addgene has a dCas9 KRAB plasmid for transient knockouts!  Turn it on ...? Check out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off control all the...

...dead Cas9 (dCas9) based activators (such as dCas9-VP64) or repressors (dCas9-KRAB or dCas9-SRDX) can also...

... 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...

...permanently modifying the genome dCas9-repressor (such as dCas9-KRAB) or dCas9 gRNA(s) targeting promoter elements... elements of target gene dCas9-KRAB is more effective than dCas9 alone for mammalian cell lines. Activate...without permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter elements...activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs should be targeted to...

... 46911 pHR-SFFV-dCas9-BFP-KRAB Human expression vector containing SFFV promoter, dCas9 that is fused to...constructs 46910 pHR-SFFV-dCas9-BFP Human expression vector containing SFFV promoter, dCas9 that is fused to 2x... to 2x NLS, tagBFP and a KRAB domain 46912 pMSCV-LTR-dCas9-VP64-BFP Human expression vector containing...MSCV LTR promoter, dCas9 that is fused to 2x NLS, VP64 and tagBFP 51023 pSLQ1658-dCas9-EGFP Human expression...46913 pMSCV-LTR-dCas9-p65AD-BFP Human expression vector containing MSCV LTR promoter, dCas9 that is fused... CXCR4 gene 46920 pTDH3-dCas9 Yeast CEN/ARS vector (Leu2) that contains dCas9 fused to NLS controlled ... promoter 46921 pTDH3-dCas9-Mxi1 Yeast CEN/ARS vector (Leu2) that contains dCas9 fused to N 51025 pSLQ1661...

...10 cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would...

...dilutions of the lentiviral vector pHAGE EF1α dCas9-KRAB . The following day, cells were treated with ...

...Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down...Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors... separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. Want more information...

...Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down...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...specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your... separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene... separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene...

...permanently modifying the genome dCas9-repressor (such as dCas9-KRAB) or dCas9 gRNA(s) targeting promoter elements... elements of target gene dCas9-KRAB is more effective than dCas9 alone for mammalian cell lines. Activate...without permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter elements...activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs should be targeted to...

...Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down...Promoter PI Publication 64325 3xFLAG-dCas9/p-bacteria 3xFLAG-dCas9 pLtetO-1 Fujii Efficient isolation of...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 ...Promoter PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can...specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your... separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene...

...achieved by using orthogonal dCas9s (e.g. S. pyogenes dCas9 and S. aureus dCas9) tagged with different fluorescent...human cells. The simplest dCas9-based activators and repressors consist of dCas9 fused directly to a single...imaging of proteins in living cells (Figure 9B) dCas9-VPR - dCas9 fused to several different activation domains...Jonathan Weissman labs , is an all-in-one dCas9 fusion with KRAB, DNMT3A, and DNMT3L. CRISPRoff maintains...utilizes biotin tagging of dCas9 by fusing a biotin acceptor site to dCas9 and co-expressing BirA biotin... defense CRISPRa CRISPR Activation; using a dCas9 or dCas9-activator with a gRNA to increase transcription...target gene CRISPRi CRISPR Interference; using a dCas9 or dCas9-repressor with a gRNA to repress/decrease transcription...

...Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down...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 ...Marker PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can...specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your... separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene... separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene...

...Catalytically dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down...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...specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your... separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene... separate gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene...