OAT2 CRISPR Activation Plasmid (h): sc-403923-ACT

SLC22A7 encodes the human organic anion transporter 2 (OAT2), a polyspecific solute carrier that mediates cellular uptake and exchange of endogenous organic anions and xenobiotic metabolites. OAT2 activity contributes to hepatic and renal handling of small molecules, influencing intracellular anion balance and coupling to metabolic and detoxification processes through coordinated transporter networks. By shaping cellular exposure to drug-like compounds and metabolic intermediates, OAT2 is relevant to studies of pharmacokinetics, transporter-mediated drug disposition, and variability in tissue-specific chemical sensitivity. Dysregulated transporter expression or function is therefore of interest in mechanistic research on liver and kidney physiology and disease-associated alterations in xenobiotic clearance pathways.

OAT2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC22A7 expression without altering the underlying DNA sequence.

OAT2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC22A7 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 SLC22A7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OAT2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC22A7 locus and enabling the study of OAT2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OAT2 pathway restoration in tumor cells with silenced or reduced SLC22A7 expression.

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