mPRα CRISPR Activation Plasmid (h): sc-402143-ACT

PAQR7 encodes membrane progesterone receptor alpha (mPRα), a member of the PAQR family implicated in rapid, non-genomic progesterone signaling at cellular membranes. mPRα has been linked to modulation of G protein–coupled signaling, including cAMP dynamics and downstream kinase pathways that influence cell proliferation, differentiation, and survival programs. In human tissues, PAQR7 expression is associated with endocrine regulation and steroid-responsive cellular states, making it relevant for studying hormone-dependent transcriptional networks and membrane-initiated signaling cross-talk. Dysregulated progesterone signaling pathways involving PAQR7 have been investigated in the context of reproductive biology and hormone-associated disease mechanisms, supporting its use as a research target in cell signaling and functional genomics studies.

mPRα CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PAQR7 expression without altering the underlying DNA sequence.

mPRα CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PAQR7 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 PAQR7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous mPRα expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PAQR7 locus and enabling the study of mPRα-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of mPRα pathway restoration in tumor cells with silenced or reduced PAQR7 expression.

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