α-SNAP CRISPR Activation Plasmid (h): sc-403996-ACT

NAPA encodes human α-SNAP, a soluble NSF attachment protein that binds SNARE complexes and cooperates with NSF to drive ATP-dependent SNARE disassembly, enabling repeated rounds of vesicle docking and membrane fusion. Through this chaperone-like function, α-SNAP supports constitutive and regulated exocytosis, endocytosis, and Golgi-to-ER trafficking, thereby influencing secretion, receptor recycling, and organelle homeostasis. Perturbation of NAPA-dependent SNARE cycling can disrupt proteostasis and membrane dynamics, processes that intersect with synaptic function and stress responses. Dysregulated vesicle trafficking and SNARE complex turnover are implicated in neurodegenerative and neurodevelopmental disease mechanisms, making α-SNAP a useful node for pathway-focused studies of intracellular transport.

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

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

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