Catalytically deactivated, or dead Cas9 (dCas9) expression is a key component in performing efficient CRISPR activation (CRISPRa) or CRISPR interference (CRISPRi) experiments. Cas9 has been modified to remove the nuclease activity but to keep the highly-specific genomic DNA targeting capability with the help of guide RNA. The deactivated Cas9 (dCas9) for CRISPRa is fused to a series of transcription activators (VP64, p65 and Rta) to create a dCas9-VPR complex which, paired with specific gRNAs, upregulates gene expression. For CRISPRi, the dCas9 is fused to transcriptional repressor peptide, KRAB, to disrupt gene expression when paired with specific gRNAs.
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