Syntaxin 3 CRISPR Activation Plasmid (h): sc-403466-ACT

STX3 encodes syntaxin 3, a plasma membrane–localized t-SNARE that mediates vesicle docking and membrane fusion during polarized exocytosis. Syntaxin 3 functions with SNAP family proteins and VAMPs to regulate apical trafficking in epithelial cells and targeted delivery of cargos in neurons and immune cells, influencing membrane protein recycling and secretion. Through its role in endosomal sorting and exocytic pathways, STX3 contributes to maintenance of cell polarity, junctional organization, and barrier function. Dysregulated SNARE-dependent trafficking involving STX3 has been implicated in disorders of epithelial polarity and neural function, supporting its study in pathways relevant to cellular organization and disease-associated transport phenotypes.

Syntaxin 3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous STX3 expression without altering the underlying DNA sequence.

Syntaxin 3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the STX3 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 STX3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Syntaxin 3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native STX3 locus and enabling the study of Syntaxin 3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Syntaxin 3 pathway restoration in tumor cells with silenced or reduced STX3 expression.

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