PGRMC2 CRISPR Activation Plasmid (h): sc-405132-ACT

Progesterone receptor membrane component 2 (PGRMC2) is a heme-binding membrane-associated protein implicated in sterol metabolism, membrane trafficking, and regulation of cytochrome P450–dependent oxidative reactions. In human cells, PGRMC2 has been linked to intracellular cholesterol handling and endoplasmic reticulum–associated processes that shape lipid homeostasis and cellular stress responses. Its activity intersects with pathways governing mitochondrial/ER function, redox balance, and protein complex organization, making it relevant to studies of metabolic reprogramming. Altered PGRMC2 expression or signaling has been associated with phenotypes observed in cancer biology, endocrine regulation, and inflammatory states, supporting its utility as a mechanistic node in disease-relevant models.

PGRMC2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PGRMC2 expression without altering the underlying DNA sequence.

PGRMC2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PGRMC2 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 PGRMC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PGRMC2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PGRMC2 locus and enabling the study of PGRMC2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PGRMC2 pathway restoration in tumor cells with silenced or reduced PGRMC2 expression.

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