VAMP-3 CRISPR Activation Plasmid (h): sc-402306-ACT

VAMP3 encodes vesicle-associated membrane protein 3 (VAMP-3), a v-SNARE that mediates membrane fusion events required for endocytic recycling and regulated exocytosis. It localizes predominantly to recycling endosomes and supports trafficking of receptors and transporters to the plasma membrane, coordinating processes such as cell migration, adhesion turnover, and cytokine or enzyme secretion. VAMP-3 participates in SNARE complex assembly with syntaxins and SNAP proteins, linking vesicular transport to cytoskeletal remodeling and immune cell function. Dysregulated vesicle trafficking involving VAMP-3 has been associated with altered inflammatory signaling, pathogen entry/egress, and metastatic phenotypes, making it relevant for studies of innate immunity, infection biology, and cancer cell invasion.

VAMP-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VAMP3 expression without altering the underlying DNA sequence.

VAMP-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VAMP3 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 VAMP3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous VAMP-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VAMP3 locus and enabling the study of VAMP-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of VAMP-3 pathway restoration in tumor cells with silenced or reduced VAMP3 expression.

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