Syntaxin 5 CRISPR Activation Plasmid (h): sc-402516-ACT

Human STX5 encodes syntaxin 5, a core Qa-SNARE that mediates vesicle docking and membrane fusion within the early secretory pathway, supporting ER-to-Golgi transport and intra-Golgi trafficking. By coordinating SNARE complex assembly at the cis-Golgi and ER–Golgi intermediate compartment, STX5 helps maintain Golgi organization, protein glycosylation, and polarized secretion. Perturbation of STX5-dependent trafficking can disrupt proteostasis and glycan processing, impacting pathways that govern receptor maturation, membrane protein localization, and secretory cargo handling. Altered secretory pathway function involving STX5 has been linked in the literature to cellular stress responses and phenotypes relevant to congenital disorders of glycosylation and broader trafficking-associated disease mechanisms.

Syntaxin 5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous STX5 expression without altering the underlying DNA sequence.

Syntaxin 5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the STX5 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 STX5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Syntaxin 5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native STX5 locus and enabling the study of Syntaxin 5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Syntaxin 5 pathway restoration in tumor cells with silenced or reduced STX5 expression.

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