ABCG8 CRISPR Activation Plasmid (h): sc-402453-ACT

ABCG8 encodes an ATP-binding cassette (ABC) half-transporter that heterodimerizes with ABCG5 to form a functional sterol efflux pump at the apical membrane of enterocytes and canalicular membrane of hepatocytes. This complex limits intestinal absorption and promotes biliary excretion of dietary sterols, shaping whole-body cholesterol and phytosterol homeostasis through coordinated transport and lipid handling pathways. Disruption of ABCG8 activity perturbs sterol trafficking and is linked to dysregulated plasma sterol profiles and inherited sterol accumulation phenotypes, including sitosterolemia-like presentations. As a result, ABCG8 is widely studied in metabolic regulation, lipid transport biology, and genotype–phenotype relationships in sterol metabolism.

ABCG8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCG8 expression without altering the underlying DNA sequence.

ABCG8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCG8 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 ABCG8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ABCG8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCG8 locus and enabling the study of ABCG8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ABCG8 pathway restoration in tumor cells with silenced or reduced ABCG8 expression.

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