ABCG5 CRISPR Activation Plasmid (m): sc-424275-ACT

Mouse Abcg5 encodes the ATP-binding cassette transporter ABCG5, which heterodimerizes with ABCG8 to mediate sterol efflux across apical membranes in enterocytes and hepatocytes. This transporter complex limits intestinal absorption of dietary sterols and promotes biliary secretion of cholesterol and plant sterols, integrating with lipid homeostasis pathways that include nuclear receptor regulation and bile acid metabolism. Disruption of ABCG5-dependent transport alters sterol trafficking and contributes to dyslipidemia-related phenotypes, making Abcg5 a relevant target for studying intestinal-liver axis control of lipid balance. In vivo and in vitro models leveraging Abcg5 modulation support mechanistic investigations of metabolic stress, hepatic lipid handling, and sterol-driven inflammatory responses.

ABCG5 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Abcg5 expression without altering the underlying DNA sequence.

ABCG5 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Abcg5 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 Abcg5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ABCG5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Abcg5 locus and enabling the study of ABCG5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ABCG5 pathway restoration in tumor cells with silenced or reduced Abcg5 expression.

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