Syntaxin 17 CRISPR Activation Plasmid (h): sc-403251-ACT

STX17 encodes Syntaxin 17, a SNARE-family protein that localizes to the endoplasmic reticulum and mitochondria-associated membranes and supports membrane fusion events central to cellular homeostasis. Syntaxin 17 is a key mediator of autophagosome–lysosome fusion, coordinating late-stage autophagy and mitophagy with the delivery of cargo to degradative compartments. Through its role in vesicular trafficking and organelle quality control, STX17 influences proteostasis, stress adaptation, and metabolic signaling pathways that intersect with mitochondrial function. Dysregulation of autophagy and mitochondrial clearance pathways in which Syntaxin 17 participates is broadly implicated in neurodegeneration, infection biology, and cancer-associated cellular stress phenotypes, motivating mechanistic studies of STX17 expression control.

Syntaxin 17 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous STX17 expression without altering the underlying DNA sequence.

Syntaxin 17 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the STX17 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 STX17 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Syntaxin 17 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native STX17 locus and enabling the study of Syntaxin 17-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Syntaxin 17 pathway restoration in tumor cells with silenced or reduced STX17 expression.

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