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Cat. No.: DC9VP-500 (for 500pmol)

Cat. No.: DC9VP-2500 (for 2500pmol)

Description

dCas9‑VP64 is a synthetic fusion protein composed of nuclease‑dead Cas9 (dCas9) and the VP64 transcriptional activation domain, representing the core component of first‑generation CRISPR activation (CRISPRa) systems. The dCas9 module carries the D10A and N863A double mutations, which completely abolish DNA cleavage activity while preserving the ability to bind target DNA sequences with high precision under sgRNA guidance.

The VP64 activation domain consists of four tandem copies of the herpes simplex virus VP16 transcriptional activation motif. VP16 is a potent eukaryotic transcriptional activator capable of efficiently recruiting endogenous transcriptional machinery.

When a sequence‑specific sgRNA directs dCas9‑VP64 to the promoter region of a target gene, the VP64 domain recruits transcriptional components such as the pre‑initiation complex, histone acetyltransferases, and Mediator complexes. This promotes RNA polymerase II assembly and transcription initiation, resulting in upregulation of the target gene. Typically, dCas9‑VP64 achieves a 2‑ to 5‑fold increase in basal gene expression, with activation efficiency influenced by sgRNA positioning, chromatin accessibility, and cell type.

To overcome the limited activation efficiency of a single sgRNA, researchers often employ multi‑sgRNA strategies—designing several sgRNAs targeting different regions of the same promoter. Through cooperative recruitment of transcriptional machinery, this approach can dramatically enhance activation potency, with optimized systems achieving up to several hundred‑fold increases in gene expression.