mPRβ CRISPR Activation Plasmid (h): sc-403374-ACT

PAQR8 encodes membrane progesterone receptor beta (mPRβ), a seven-transmembrane progestin receptor in the PAQR family that mediates rapid, non-genomic progesterone signaling at the plasma membrane. mPRβ has been linked to regulation of second messenger pathways such as Gi/o-coupled modulation of cAMP and downstream kinase signaling, influencing cell cycle control, differentiation, and stress-responsive transcriptional programs. In human tissues, PAQR8 expression has been studied in reproductive and endocrine contexts where progesterone-dependent signaling intersects with steroid hormone networks and membrane receptor cross-talk. Dysregulated progesterone signaling and altered PAQR family receptor expression are frequently evaluated in hormone-responsive biology, including models of reproductive disorders and cancer-associated pathway remodeling, supporting PAQR8 as a target for mechanistic studies of steroid signaling dynamics.

mPRβ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PAQR8 expression without altering the underlying DNA sequence.

mPRβ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PAQR8 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 PAQR8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous mPRβ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PAQR8 locus and enabling the study of mPRβ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of mPRβ pathway restoration in tumor cells with silenced or reduced PAQR8 expression.

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