SNAP 23 CRISPR Activation Plasmid (h): sc-417781-ACT

SNAP23 encodes synaptosomal-associated protein 23, a Qbc-SNARE that pairs with syntaxins and VAMP family members to catalyze membrane fusion events required for regulated and constitutive exocytosis. SNAP23 supports vesicle docking and trafficking at the plasma membrane, influencing secretion, endocytic recycling, and GLUT4 translocation in insulin-responsive cells. Through control of granule and vesicle fusion, it contributes to immune cell degranulation and cytokine release, and its dysregulation has been associated with altered inflammatory signaling and metabolic phenotypes relevant to diabetes and obesity research. SNAP23 also interfaces with pathways governing membrane dynamics and cell surface receptor availability, linking vesicle trafficking to signaling output in diverse human cell types.

SNAP 23 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNAP23 expression without altering the underlying DNA sequence.

SNAP 23 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNAP23 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 SNAP23 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SNAP 23 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNAP23 locus and enabling the study of SNAP 23-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SNAP 23 pathway restoration in tumor cells with silenced or reduced SNAP23 expression.

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