COPG CRISPR Activation Plasmid (h): sc-404407-ACT

COPG1 encodes the COPG subunit of the COPI coatomer complex, a key mediator of vesicular transport between the Golgi apparatus and endoplasmic reticulum and within Golgi cisternae. COPG participates in coat assembly and cargo selection required for retrograde trafficking, maintenance of Golgi organization, and proteostasis, linking it to cellular stress responses and secretory pathway homeostasis. Perturbation of COPI-dependent trafficking can alter receptor recycling, lipid handling, and glycosylation, processes that influence cell growth and signaling networks. Dysregulated vesicle transport and Golgi/ER dynamics are frequently implicated in cancer biology, neurodegeneration, and immune cell function, making COPG1 a relevant target for mechanistic studies of membrane trafficking pathways.

COPG CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COPG1 expression without altering the underlying DNA sequence.

COPG CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COPG1 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 COPG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COPG expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COPG1 locus and enabling the study of COPG-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COPG pathway restoration in tumor cells with silenced or reduced COPG1 expression.

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