Vav CRISPR Activation Plasmid (h): sc-400606-ACT

Human VAV1 encodes Vav, a hematopoietic-specific guanine nucleotide exchange factor that activates Rho family GTPases such as RAC1 to coordinate actin cytoskeletal remodeling, cell adhesion, and migration. Vav functions downstream of immunoreceptor and integrin signaling, linking tyrosine kinase–dependent phosphorylation events to pathways including T cell receptor signaling, NF-κB/MAPK activation, and immune synapse formation. Dysregulated VAV1 signaling is associated with altered lymphocyte development and aberrant immune activation, and has been implicated in hematologic malignancies and immune-related disorders. As a signaling hub, VAV1 is widely studied for its roles in lymphocyte activation thresholds, cytokine production, and cytoskeleton-dependent trafficking.

Vav CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VAV1 expression without altering the underlying DNA sequence.

Vav CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VAV1 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 VAV1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Vav expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VAV1 locus and enabling the study of Vav-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Vav pathway restoration in tumor cells with silenced or reduced VAV1 expression.

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