Apolipoprotein A I/APOA1 CRISPR Activation Plasmid (m): sc-419158-ACT

Mouse Apoa1 encodes apolipoprotein A-I (APOA1), the principal protein component of high-density lipoprotein that drives cholesterol and phospholipid efflux and supports reverse cholesterol transport. APOA1 functions with lipid transporters and enzymes such as ABCA1/ABCG1 and LCAT to promote nascent HDL biogenesis, lipid remodeling, and systemic lipid homeostasis, linking hepatic secretion to peripheral cholesterol clearance. Through these processes, Apoa1 activity intersects with macrophage foam cell biology, inflammatory signaling, and oxidative stress responses in metabolic tissues. Altered Apoa1/APOA1 expression is widely used as a molecular readout in studies of dyslipidemia, atherosclerosis susceptibility, and cardiometabolic trait regulation in mouse models.

Apolipoprotein A I/APOA1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Apoa1 expression without altering the underlying DNA sequence.

Apolipoprotein A I/APOA1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Apoa1 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 Apoa1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Apolipoprotein A I/APOA1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Apoa1 locus and enabling the study of Apolipoprotein A I/APOA1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Apolipoprotein A I/APOA1 pathway restoration in tumor cells with silenced or reduced Apoa1 expression.

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