CNP-53 CRISPR Activation Plasmid (h): sc-403527-ACT

NPPC encodes C-type natriuretic peptide (CNP-53), a secreted peptide hormone that signals primarily through natriuretic peptide receptor 2 (NPR2/GC-B) to elevate intracellular cGMP and activate PKG-dependent pathways. In human tissues, CNP-53 regulates endochondral ossification, chondrocyte proliferation and differentiation, and vascular homeostasis, integrating with cGMP signaling to modulate extracellular matrix remodeling and cellular growth programs. NPPC–NPR2 signaling intersects with MAPK and calcium-regulated processes that shape cartilage development and skeletal growth. Genetic disruption of NPPC or its receptor axis is associated with disordered longitudinal bone growth and skeletal dysplasias, making this pathway relevant for modeling growth plate biology and connective tissue phenotypes.

CNP-53 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NPPC expression without altering the underlying DNA sequence.

CNP-53 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NPPC 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 NPPC transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CNP-53 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NPPC locus and enabling the study of CNP-53-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CNP-53 pathway restoration in tumor cells with silenced or reduced NPPC expression.

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