NPR-A CRISPR Activation Plasmid (h): sc-401737-ACT

Human NPR1 encodes natriuretic peptide receptor A (NPR-A), a single-pass transmembrane guanylyl cyclase that binds atrial and brain natriuretic peptides to generate cGMP. NPR-A signaling activates cGMP-dependent effectors such as PKG and modulates ion transport, vasodilatory tone, and transcriptional programs that regulate cardiovascular and renal homeostasis. Through cGMP turnover and cross-talk with nitric oxide and phosphodiesterase pathways, NPR1 influences cellular growth responses and extracellular matrix remodeling. Altered NPR1/NPR-A activity has been associated with dysregulated blood pressure control, cardiac hypertrophy, and cardio-renal pathophysiology, supporting its relevance for mechanistic studies of cardiometabolic signaling.

NPR-A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NPR1 expression without altering the underlying DNA sequence.

NPR-A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NPR1 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 NPR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NPR-A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NPR1 locus and enabling the study of NPR-A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NPR-A pathway restoration in tumor cells with silenced or reduced NPR1 expression.

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