WDR77 CRISPR Activation Plasmid (h): sc-403890-ACT

WDR77 (also known as MEP50) encodes a WD-repeat scaffold protein that functions as an essential cofactor for PRMT5, coordinating symmetric dimethylation of arginine residues on histones and diverse RNA-binding proteins. Through this PRMT5–WDR77 complex, WDR77 contributes to chromatin remodeling, transcriptional regulation, and RNA splicing programs that govern cell cycle progression and lineage-specific gene expression. WDR77-associated methylation networks intersect with pathways controlling DNA damage responses and signal-dependent transcription, linking epigenetic regulation to cellular proliferation states. Dysregulation of PRMT5/WDR77 activity and downstream methylation signatures has been reported across multiple cancer and developmental contexts, motivating mechanistic studies of WDR77-dependent gene control.

WDR77 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous WDR77 expression without altering the underlying DNA sequence.

WDR77 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the WDR77 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the WDR77 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous WDR77 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native WDR77 locus and enabling the study of WDR77-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of WDR77 pathway restoration in tumor cells with silenced or reduced WDR77 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.