MRP5 CRISPR Activation Plasmid (h): sc-402443-ACT

ABCC5 encodes the human ATP-binding cassette transporter MRP5 (ABCC5), a plasma membrane efflux pump that exports cyclic nucleotides such as cGMP and cAMP as well as select nucleoside analogs and organic anions. By controlling intracellular cyclic nucleotide pools, MRP5 can influence cGMP/PKG and cAMP/PKA signaling, with downstream effects on vascular tone regulation, platelet reactivity, and cellular stress responses. ABCC5 expression and transporter activity are frequently examined in the context of xenobiotic handling and multidrug resistance phenotypes, where altered efflux capacity can reshape intracellular metabolite and drug analog exposure. Dysregulated ABCC5 has been associated with disease-relevant changes in signaling and transport homeostasis, making it a useful target for mechanistic studies of transporter-mediated regulation.

MRP5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCC5 expression without altering the underlying DNA sequence.

MRP5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCC5 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 ABCC5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRP5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCC5 locus and enabling the study of MRP5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRP5 pathway restoration in tumor cells with silenced or reduced ABCC5 expression.

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