MRP4 CRISPR Activation Plasmid (h): sc-401912-ACT

Human ABCC4 encodes MRP4 (multidrug resistance-associated protein 4), an ATP-binding cassette transporter that mediates ATP-dependent efflux of organic anions including cyclic nucleotides (cAMP/cGMP), prostaglandins, leukotrienes, bile acid conjugates, and diverse xenobiotic metabolites. By regulating intracellular second messengers and lipid mediators, MRP4 influences GPCR signaling, inflammatory and vascular homeostasis pathways, and cellular responses to stress. ABCC4 activity also shapes pharmacokinetics and cellular exposure to nucleoside analogs and other small molecules through transport-driven clearance. Dysregulated ABCC4/MRP4 expression has been associated with altered inflammatory signaling and transporter-linked drug response phenotypes in multiple disease contexts, making it a useful target for mechanistic studies of transporter biology and signaling crosstalk.

MRP4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCC4 expression without altering the underlying DNA sequence.

MRP4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCC4 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 ABCC4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRP4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCC4 locus and enabling the study of MRP4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRP4 pathway restoration in tumor cells with silenced or reduced ABCC4 expression.

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