apoA-II CRISPR Activation Plasmid (h): sc-404987-ACT

Human APOA2 encodes apolipoprotein A-II (apoA-II), a major structural component of high-density lipoprotein (HDL) that modulates lipoprotein particle stability, lipid binding, and exchange with plasma enzymes and transfer proteins. apoA-II influences cholesterol and triglyceride homeostasis by shaping HDL remodeling and affecting interactions with pathways that govern lipoprotein metabolism, including lecithin–cholesterol acyltransferase activity and lipid transfer processes. Altered APOA2 expression or apoA-II levels have been associated with dyslipidemia-related phenotypes and cardiometabolic risk traits, supporting its relevance for mechanistic studies in lipid biology. In hepatic and intestinal contexts, apoA-II contributes to systemic lipid transport and can impact inflammatory and metabolic signaling linked to atherosclerosis-associated processes.

apoA-II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous APOA2 expression without altering the underlying DNA sequence.

apoA-II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the APOA2 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 APOA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous apoA-II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native APOA2 locus and enabling the study of apoA-II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of apoA-II pathway restoration in tumor cells with silenced or reduced APOA2 expression.

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