...repressor (such as dCas9-KRAB) or dCas9 gRNA(s) targeting promoter elements of target gene dCas9-KRAB is more ...permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter elements...CRISPRi) Reduce expression of a particular gene(s) without permanently modifying the genome dCas9-repressor ...effective than dCas9 alone for mammalian cell lines. Activate (CRISPRa) Increase expression of an endogenous...activators exist, including the multi-plasmid SAM system. Select Expression System To use CRISPR, you will need...to the transient Cas9 expression. The table below summarizes the major expression systems and variables...Delivery method Expression System Components of System Application Mammalian expression vector Cas enzyme...

...system (Figure 9C) SAM activators - co-expression of dCas9-VP64 with a modified scaffold gRNA and additional...tools require constitutive expression of the dCas9 fusion and/or expression of multiple fusions. The effects...achieved by using orthogonal dCas9s (e.g. S. pyogenes dCas9 and S. aureus dCas9) tagged with different fluorescent...biotin tagging of dCas9 by fusing a biotin acceptor site to dCas9 and co-expressing BirA biotin ligase...multiple gRNAs using a single plasmid ensures that all gRNAs will be expressed in the same cell. This is ...genomic DNA. dCas9 has also been used for genome-wide screens to activate or repress gene expression in mice...human cells. The simplest dCas9-based activators and repressors consist of dCas9 fused directly to a single...