GS27 CRISPR Activation Plasmid (h): sc-406352-ACT

Human GOSR2 encodes GS27, a Qb-SNARE localized to the cis-Golgi and ER–Golgi intermediate compartment that supports vesicle tethering and membrane fusion within the early secretory pathway. By partnering with other SNAREs such as syntaxin 5, BET1, and SEC22 family members, GS27 helps maintain Golgi organization, cargo trafficking, and protein processing through COPI/COPII-dependent transport cycles. Disruption of GOSR2 function has been linked to neurodevelopmental phenotypes and progressive myoclonus epilepsy, consistent with the sensitivity of neurons to secretory pathway stress and proteostasis imbalance. As a result, GS27 is frequently studied in contexts including ER stress responses, glycoprotein maturation, and Golgi-to-ER retrograde trafficking.

GS27 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GOSR2 expression without altering the underlying DNA sequence.

GS27 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GOSR2 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 GOSR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GS27 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GOSR2 locus and enabling the study of GS27-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GS27 pathway restoration in tumor cells with silenced or reduced GOSR2 expression.

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