Exo84 CRISPR Activation Plasmid (h): sc-403472-ACT

Human EXOC8 encodes Exo84, a core component of the octameric exocyst tethering complex that directs secretory vesicles to defined sites on the plasma membrane prior to SNARE-mediated fusion. Through its role in polarized exocytosis, Exo84 supports processes including epithelial polarity establishment, neurite outgrowth, ciliogenesis, and directed cell migration, and it interfaces with small GTPase signaling (for example Ral and Rab pathways) that coordinates vesicle targeting and cytoskeletal remodeling. Exocyst-dependent trafficking also influences surface delivery and recycling of receptors and transporters, shaping signaling outputs and membrane composition. Dysregulated exocyst function has been associated with altered cell polarity and invasive behavior in cancer models, as well as defects in neuronal and epithelial homeostasis linked to developmental and neurodegenerative phenotypes.

Exo84 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous EXOC8 expression without altering the underlying DNA sequence.

Exo84 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the EXOC8 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 EXOC8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Exo84 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native EXOC8 locus and enabling the study of Exo84-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Exo84 pathway restoration in tumor cells with silenced or reduced EXOC8 expression.

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