BNP CRISPR Activation Plasmid (h): sc-402447-ACT

Human NPPB encodes B-type natriuretic peptide (BNP), a cardiac-derived hormone produced primarily by ventricular cardiomyocytes in response to increased wall stress. Following prohormone processing, BNP signals through natriuretic peptide receptor A (NPR1/GC-A) to elevate cGMP and activate PKG-dependent pathways that modulate vasodilation, natriuresis, and anti-fibrotic gene programs. NPPB transcription integrates biomechanical and neurohumoral inputs, including calcium-dependent signaling and MAPK cascades, linking hemodynamic load to coordinated changes in myocardial remodeling. Dysregulated NPPB/BNP expression is widely used as a molecular indicator of cardiac stress and is studied in contexts such as hypertrophy, fibrosis, and heart failure biology.

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

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

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