SNAP 25 CRISPR Activation Plasmid (h): sc-401138-ACT

Human SNAP25 encodes synaptosomal-associated protein 25, a core t-SNARE that assembles with syntaxin and VAMP/synaptobrevin to drive synaptic vesicle docking and Ca²⁺-triggered membrane fusion. SNAP 25 activity is central to regulated exocytosis, shaping neurotransmitter release probability, short-term plasticity, and presynaptic vesicle recycling within the SNARE-mediated vesicle trafficking pathway. By coupling vesicle fusion to calcium signaling and presynaptic cytoskeletal organization, SNAP 25 influences neuronal network excitability and circuit maturation. Altered SNAP25 expression or function has been linked in genetic and functional studies to neurodevelopmental and neuropsychiatric phenotypes, making it a useful node for investigating synaptic dysfunction mechanisms.

SNAP 25 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SNAP25 expression without altering the underlying DNA sequence.

SNAP 25 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SNAP25 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 SNAP25 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SNAP 25 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SNAP25 locus and enabling the study of SNAP 25-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SNAP 25 pathway restoration in tumor cells with silenced or reduced SNAP25 expression.

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