...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...