Vav2 CRISPR Activation Plasmid (h): sc-401490-ACT

Human VAV2 encodes Vav2, a Rho family guanine nucleotide exchange factor that catalyzes GDP–GTP exchange on Rac1, RhoA, and Cdc42 to coordinate actin cytoskeleton remodeling, cell adhesion, and directional migration. Vav2 functions downstream of receptor tyrosine kinases, integrins, and immune receptors, linking extracellular cues to cytoskeletal and transcriptional programs through pathways that intersect with MAPK and PI3K signaling. Through these roles, Vav2 helps regulate cell morphology, endocytosis, and signal-dependent changes in motility and invasion-related phenotypes. Dysregulated VAV2 expression or signaling has been associated with aberrant proliferation and migration programs in cancer biology and with altered immune and vascular signaling contexts relevant to disease mechanisms.

Vav2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VAV2 expression without altering the underlying DNA sequence.

Vav2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VAV2 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 VAV2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Vav2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VAV2 locus and enabling the study of Vav2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Vav2 pathway restoration in tumor cells with silenced or reduced VAV2 expression.

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