ABCG2 CRISPR Activation Plasmid (h): sc-416737-ACT

ABCG2 (ATP-binding cassette sub-family G member 2) is a plasma membrane efflux transporter that uses ATP hydrolysis to export a broad range of endogenous metabolites and xenobiotics. It is highly expressed in barrier tissues and in stem-like cell compartments, where it contributes to the “side population” phenotype and regulates intracellular accumulation of small molecules. ABCG2 activity intersects with detoxification and oxidative stress–related processes, influencing cellular homeostasis under metabolic and environmental challenge. Altered ABCG2 expression or function is associated with variability in drug disposition and has been studied in cancer biology and tissue protection contexts where transporter-mediated efflux shapes cellular sensitivity to chemical stressors.

ABCG2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCG2 expression without altering the underlying DNA sequence.

ABCG2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCG2 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 ABCG2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ABCG2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCG2 locus and enabling the study of ABCG2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ABCG2 pathway restoration in tumor cells with silenced or reduced ABCG2 expression.

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