MATE2 CRISPR Activation Plasmid (h): sc-404811-ACT

Human SLC47A2 encodes the multidrug and toxin extrusion transporter MATE2 (SLC47A2), an electroneutral organic cation/H+ antiporter that mediates efflux of endogenous metabolites and xenobiotic cations across epithelial membranes. MATE2 is most prominently linked to renal proximal tubule transport, where it functions in concert with uptake transporters such as OCT2 to coordinate vectorial secretion of organic cations and shape cellular exposure to charged compounds. By controlling intracellular accumulation of cationic substrates, SLC47A2 influences pharmacokinetic variability, transporter–transporter interactions, and stress responses associated with altered epithelial transport capacity. Dysregulated expression or functional variation in SLC47A2 is therefore relevant to studies of kidney physiology, drug disposition pathways, and transporter-associated susceptibility to renal injury phenotypes in experimental models.

MATE2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC47A2 expression without altering the underlying DNA sequence.

MATE2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC47A2 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 SLC47A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MATE2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC47A2 locus and enabling the study of MATE2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MATE2 pathway restoration in tumor cells with silenced or reduced SLC47A2 expression.

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