MRP3 CRISPR Activation Plasmid (h): sc-401508-ACT

ABCC3 encodes the human ATP-binding cassette transporter MRP3, a basolateral efflux pump that exports a broad range of organic anions, including glucuronide and sulfate conjugates and bile acids. MRP3 supports hepatic detoxification and enterohepatic circulation by coordinating metabolite clearance and modulating intracellular exposure to xenobiotics, linking transporter activity to phase II metabolism and cellular stress responses. Its expression is regulated in contexts of cholestasis and inflammatory signaling, and altered ABCC3/MRP3 activity has been associated with variability in drug disposition and hepatobiliary dysfunction in experimental models. These properties make ABCC3 a useful target for studying transporter-mediated barriers, metabolic rewiring, and adaptive responses in liver and epithelial systems.

MRP3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCC3 expression without altering the underlying DNA sequence.

MRP3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCC3 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 ABCC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRP3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCC3 locus and enabling the study of MRP3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRP3 pathway restoration in tumor cells with silenced or reduced ABCC3 expression.

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