Sec8 CRISPR Activation Plasmid (h): sc-405009-ACT

Human EXOC4 encodes Sec8, a core component of the octameric exocyst complex that tethers post-Golgi secretory vesicles to the plasma membrane prior to SNARE-dependent membrane fusion. Sec8 supports polarized exocytosis, membrane protein delivery, and cytoskeletal coordination, linking vesicle trafficking with cell migration, neurite outgrowth, and epithelial polarity programs. Through exocyst-dependent trafficking, EXOC4 influences signaling and receptor surface availability, with downstream effects on growth factor responsiveness and cell–cell junction maintenance. Dysregulation of exocyst function, including altered EXOC4/Sec8 activity, has been associated with defects in secretion and polarity that are frequently studied in the context of invasive behavior and other complex disease-relevant cellular phenotypes.

Sec8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EXOC4 expression without altering the underlying DNA sequence.

Sec8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EXOC4 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 EXOC4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sec8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EXOC4 locus and enabling the study of Sec8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sec8 pathway restoration in tumor cells with silenced or reduced EXOC4 expression.

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