dCas9-GFP-VP64 Transcriptional Activator

Product Introduction

dCas9-GFP-VP64 is a multifunctional chimeric protein that fuses nuclease-dead Cas9 (dCas9), Green Fluorescent Protein (GFP), and the VP64 transcriptional activation domain into a single entity, enabling the integration of gene targeting, real-time visualization, and transcriptional regulation. The dCas9 moiety of this protein carries the D10A and N863A double mutations, completely abolishing DNA cleavage activity while fully retaining the ability to precisely bind specific DNA sequences guided by sgRNA. GFP serves as a fluorescent reporter tag; its stable folding and bright fluorescence provide a convenient optical tool for real-time tracking of the protein's subcellular localization, dynamic distribution, and target binding process. The VP64 domain consists of four tandem copies of the herpes simplex virus VP16 transcriptional activation domain, possessing the potent ability to recruit transcriptional machinery.

When a specific sgRNA guides dCas9-GFP-VP64 to bind to the promoter region of a target gene, the VP64 domain recruits factors such as the pre-initiation complex and histone acetyltransferases, promoting the assembly of RNA polymerase II and transcription initiation, thereby achieving transcriptional upregulation of the target gene.