PGRMC1 CRISPR Activation Plasmid (h): sc-401945-ACT

Progesterone receptor membrane component 1 (PGRMC1) is a heme-binding membrane-associated protein implicated in steroid signaling, cytochrome P450 regulation, and cellular responses to oxidative and metabolic stress. In human cells, PGRMC1 has been linked to modulation of lipid and cholesterol homeostasis, endomembrane trafficking, and mitochondrial-associated processes that influence proliferation and survival programs. It participates in pathways affecting receptor crosstalk and kinase signaling, with reported connections to ER stress responses and redox balance. Dysregulated PGRMC1 expression has been observed across multiple disease-relevant contexts, including cancer biology, neurobiology, and metabolic phenotypes, making it a useful target for mechanistic studies of membrane signaling and cellular stress adaptation.

PGRMC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PGRMC1 expression without altering the underlying DNA sequence.

PGRMC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PGRMC1 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 PGRMC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PGRMC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PGRMC1 locus and enabling the study of PGRMC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PGRMC1 pathway restoration in tumor cells with silenced or reduced PGRMC1 expression.

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