GCC2 CRISPR Activation Plasmid (h): sc-411304-ACT

Human GCC2 (GRIP and coiled-coil domain containing 2) encodes a peripheral Golgi protein that localizes to the trans-Golgi network, where it participates in vesicle tethering and membrane trafficking important for cargo sorting and maintenance of Golgi architecture. Through interactions with small GTPase-regulated trafficking machinery, GCC2 contributes to endosome–Golgi transport dynamics that shape protein processing and secretion. Perturbation of Golgi tethering and trafficking pathways is broadly relevant to cellular homeostasis, including stress responses, polarity, and regulated secretion programs. Altered expression or function of Golgi-associated coiled-coil tethers such as GCC2 has been examined in the context of diseases where membrane trafficking and secretion are dysregulated, including cancer and neurodegeneration-related processes.

GCC2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GCC2 expression without altering the underlying DNA sequence.

GCC2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GCC2 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 GCC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GCC2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GCC2 locus and enabling the study of GCC2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GCC2 pathway restoration in tumor cells with silenced or reduced GCC2 expression.

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