ABC1 CRISPR Activation Plasmid (h): sc-401086-ACT

Human ABCA1 (ABC1) encodes an ATP-binding cassette transporter that exports cholesterol and phospholipids to lipid-poor apolipoproteins, initiating nascent HDL formation and supporting reverse cholesterol transport. ABC1 activity is integrated with LXR/RXR-regulated lipid homeostasis and influences membrane composition, lipid raft organization, and inflammatory signaling in macrophages and other cell types. Dysregulated ABCA1 expression or function perturbs cellular cholesterol balance and has been associated with atherosclerosis-related phenotypes, neurodegenerative processes, and altered immune cell responses. As a central node in cholesterol efflux pathways, ABCA1 is widely studied in lipoprotein biology, cardiometabolic research, and mechanisms linking lipid metabolism to inflammation.

ABC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCA1 expression without altering the underlying DNA sequence.

ABC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCA1 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 ABCA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ABC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCA1 locus and enabling the study of ABC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ABC1 pathway restoration in tumor cells with silenced or reduced ABCA1 expression.

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