BRWD1 CRISPR Activation Plasmid (m): sc-430062-ACT

Brwd1 encodes the mouse bromodomain and WD repeat domain-containing protein BRWD1, a nuclear chromatin-associated factor that recognizes acetylated histones and supports epigenetic regulation of transcription. BRWD1 has established roles in germ cell development and meiotic progression, and it contributes to lineage-specific gene expression programs in immune cells, including B cell differentiation. Through its bromodomains and WD repeats, BRWD1 is linked to chromatin remodeling, transcriptional control, and cell-cycle–coordinated gene regulation. Dysregulation of BRWD1-associated chromatin states is relevant to studies of developmental defects, infertility phenotypes, and immune system abnormalities in mouse models.

BRWD1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Brwd1 expression without altering the underlying DNA sequence.

BRWD1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Brwd1 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 Brwd1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BRWD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Brwd1 locus and enabling the study of BRWD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BRWD1 pathway restoration in tumor cells with silenced or reduced Brwd1 expression.

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