apoC-III CRISPR Activation Plasmid (h): sc-403887-ACT

Human APOC3 encodes apolipoprotein C-III (apoC-III), a secreted apolipoprotein that modulates triglyceride-rich lipoprotein metabolism by limiting lipoprotein lipase–mediated hydrolysis and delaying hepatic clearance of remnant particles. Through its effects on VLDL and chylomicron remnant trafficking, apoC-III influences lipid transport pathways and systemic energy homeostasis. Altered APOC3 expression is associated with dyslipidemia phenotypes characterized by elevated plasma triglycerides and changes in atherogenic lipoprotein profiles, supporting its use as a mechanistic node for studying cardiometabolic risk biology. In cellular models, APOC3 regulation can be examined alongside transcriptional control of hepatic lipid handling programs and secretory pathway processing of apolipoproteins.

apoC-III CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous APOC3 expression without altering the underlying DNA sequence.

apoC-III CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the APOC3 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 APOC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous apoC-III expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native APOC3 locus and enabling the study of apoC-III-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of apoC-III pathway restoration in tumor cells with silenced or reduced APOC3 expression.

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