MRP6 CRISPR Activation Plasmid (h): sc-402219-ACT

ABCC6 encodes the ATP-binding cassette transporter MRP6, a membrane efflux pump predominantly expressed in liver and kidney that regulates extracellular nucleotide and metabolite homeostasis. Through control of ATP release and subsequent extracellular pyrophosphate generation, MRP6 contributes to mineralization balance and connective tissue integrity, linking transporter activity to purinergic signaling and ectonucleotidase pathways. Loss-of-function or reduced expression of ABCC6 is strongly associated with ectopic calcification phenotypes, including pseudoxanthoma elasticum and related heritable mineralization disorders. These biology-driven associations make ABCC6 a valuable target for studying systemic regulators of calcification, hepatic transport networks, and extracellular matrix remodeling mechanisms.

MRP6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCC6 expression without altering the underlying DNA sequence.

MRP6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCC6 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 ABCC6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MRP6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCC6 locus and enabling the study of MRP6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MRP6 pathway restoration in tumor cells with silenced or reduced ABCC6 expression.

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