NARC-1 CRISPR Activation Plasmid (h): sc-402445-ACT

PCSK9 encodes the secreted serine protease NARC-1, a key post-translational regulator of cholesterol homeostasis through control of LDL receptor abundance at the hepatocyte surface. By promoting internalization and lysosomal degradation of LDLR, PCSK9 modulates receptor recycling and influences lipoprotein uptake, integrating with SREBP-driven lipid metabolic programs. PCSK9 expression and activity intersect with ER-to-Golgi trafficking, secretory pathway processing, and extracellular receptor–ligand dynamics. Dysregulated PCSK9 signaling is strongly associated with altered plasma LDL cholesterol and cardiometabolic disease susceptibility, making it a widely used target for mechanistic studies of lipid regulation.

NARC-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PCSK9 expression without altering the underlying DNA sequence.

NARC-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PCSK9 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 PCSK9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NARC-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PCSK9 locus and enabling the study of NARC-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NARC-1 pathway restoration in tumor cells with silenced or reduced PCSK9 expression.

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