NPR-B CRISPR Activation Plasmid (h): sc-401861-ACT

NPR2 encodes natriuretic peptide receptor B (NPR-B), a membrane guanylyl cyclase that binds C-type natriuretic peptide to generate cGMP and activate downstream PKG-dependent signaling. This pathway regulates endochondral ossification, chondrocyte proliferation and differentiation, and broader control of cellular growth programs through cGMP-mediated modulation of ion transport and transcriptional responses. NPR2 signaling intersects with skeletal development networks and growth plate homeostasis, and genetic or functional perturbation is linked to disorders of linear growth and cartilage biology. In addition to developmental phenotypes, altered natriuretic peptide–cGMP signaling is relevant to studies of vascular tone regulation and tissue remodeling in human cell models.

NPR-B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NPR2 expression without altering the underlying DNA sequence.

NPR-B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NPR2 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 NPR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NPR-B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NPR2 locus and enabling the study of NPR-B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NPR-B pathway restoration in tumor cells with silenced or reduced NPR2 expression.

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