MRP2 CRISPR Activation Plasmid (h): sc-400601-ACT

ABCC2 encodes multidrug resistance-associated protein 2 (MRP2), an ATP-binding cassette efflux transporter localized primarily to the apical membrane of polarized epithelial cells such as hepatocytes, enterocytes, and renal proximal tubule cells. MRP2 exports a broad range of endogenous and xenobiotic anions, including glutathione, glucuronide, and sulfate conjugates, supporting phase II detoxification and bile formation within hepatobiliary transport pathways. By controlling cellular efflux of conjugated metabolites, MRP2 contributes to oxidative stress handling and chemical homeostasis across barrier tissues. Altered ABCC2 expression or function has been linked to impaired biliary excretion phenotypes and variability in disposition of conjugated compounds, making it a relevant target for mechanistic studies in hepatic and epithelial biology.

MRP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCC2 expression without altering the underlying DNA sequence.

MRP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCC2 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 ABCC2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCC2 locus and enabling the study of MRP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRP2 pathway restoration in tumor cells with silenced or reduced ABCC2 expression.

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