VASP CRISPR Activation Plasmid (h): sc-401104-ACT

Human VASP (vasodilator-stimulated phosphoprotein) is an Ena/VASP family actin regulatory protein that coordinates actin filament elongation, focal adhesion dynamics, and membrane protrusion during cell spreading and motility. Through interactions with profilin-actin and F-actin, VASP integrates signals downstream of cAMP/cGMP-dependent kinases and Rho-family GTPase pathways to tune cytoskeletal remodeling and cell–cell or cell–matrix adhesion. VASP-dependent control of endothelial barrier function, platelet activation, and directed migration links this node to processes relevant to vascular biology, inflammation, and invasive cell behavior. Altered VASP activity or expression has been studied in contexts such as cardiovascular remodeling and cancer-associated migration, making it a useful handle for probing actin-driven phenotypes.

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

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

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