BRWD3 CRISPR Activation Plasmid (h): sc-415190-ACT

Human BRWD3 encodes a bromodomain and WD repeat–containing chromatin-associated protein that participates in epigenetic regulation of transcription by coupling acetyl-lysine recognition to chromatin remodeling and DNA-templated processes. Through its bromodomains and scaffolding WD repeats, BRWD3 is implicated in controlling gene expression programs that influence cell proliferation, differentiation, and genome stability. Genetic and functional studies have linked BRWD3 dysregulation to neurodevelopmental phenotypes, including X-linked intellectual disability, and to broader transcriptional perturbations relevant to cancer biology. BRWD3 therefore serves as a useful node for investigating chromatin-dependent pathway control and genotype-to-phenotype relationships in human cells.

BRWD3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BRWD3 expression without altering the underlying DNA sequence.

BRWD3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BRWD3 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 BRWD3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BRWD3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BRWD3 locus and enabling the study of BRWD3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BRWD3 pathway restoration in tumor cells with silenced or reduced BRWD3 expression.

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