BVES CRISPR Activation Plasmid (h): sc-403698-ACT

BVES (blood vessel epicardial substance), also known as POPDC1, encodes a membrane-associated protein enriched in cardiac and epithelial tissues that functions as a cAMP-binding regulator of cell–cell adhesion and membrane trafficking. BVES participates in junctional organization and cytoskeletal remodeling, influencing epithelial integrity, migration, and electrical conduction phenotypes in striated muscle. Through modulation of cAMP-dependent signaling and associated scaffolding interactions, BVES contributes to tissue homeostasis and stress responses. Altered BVES expression or localization has been linked to defects in epithelial barrier function and cardiac conduction-related phenotypes, supporting its relevance in mechanistic studies of cardiomyopathy- and epithelial dysregulation-associated pathways.

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

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

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