MATE1 CRISPR Activation Plasmid (h): sc-403332-ACT

Human SLC47A1 encodes the multidrug and toxin extrusion transporter 1 (MATE1), a polyspecific SLC transporter that functions as an electroneutral H+/organic cation antiporter on epithelial membranes. MATE1 cooperates with uptake transporters such as OCT family members to mediate vectorial transport and cellular clearance of endogenous metabolites and xenobiotic organic cations, supporting renal and hepatobiliary excretory pathways. By shaping intracellular exposure to cationic compounds, MATE1 influences cellular stress responses and transporter-regulated metabolic homeostasis. Altered SLC47A1 activity or expression is studied in the context of inter-individual variability in compound handling and transporter-associated susceptibility to tissue injury in kidney and liver model systems.

MATE1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC47A1 expression without altering the underlying DNA sequence.

MATE1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC47A1 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 SLC47A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MATE1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC47A1 locus and enabling the study of MATE1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MATE1 pathway restoration in tumor cells with silenced or reduced SLC47A1 expression.

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