BRAP CRISPR Activation Plasmid (h): sc-403847-ACT

BRAP (BRCA1-associated protein) is a cytoplasmic RING-type E3 ubiquitin ligase that modulates protein stability and signaling output by coordinating ubiquitination-dependent control of pathway components. It has been implicated in regulating MAPK/ERK signaling dynamics, influencing cell-cycle progression, stress responses, and proteostasis through effects on ubiquitin-mediated turnover and signal attenuation. By shaping signal transduction and inflammatory/stress pathways, BRAP expression and functional variation have been associated with altered cellular homeostasis and susceptibility to disease-relevant phenotypes, including cardiometabolic and neurodegeneration-linked processes. These properties make BRAP a useful target for dissecting how ubiquitin ligase networks tune phosphorylation-driven signaling and downstream transcriptional programs in human cells.

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

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

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