OAT1 CRISPR Activation Plasmid (h): sc-404284-ACT

Human SLC22A6 encodes organic anion transporter 1 (OAT1), a basolateral renal proximal tubule transporter that mediates uptake of endogenous metabolites and xenobiotic organic anions from blood into epithelial cells. OAT1 functions within solute carrier networks that couple organic anion exchange to cellular handling of urate, uremic toxins, and drug conjugates, shaping renal secretion and systemic clearance. By influencing intracellular exposure to metabolites and pharmaceuticals, SLC22A6 activity intersects with pathways linked to tubular stress responses and transport-dependent toxicology. Altered OAT1 expression or function has been associated with variation in drug disposition and susceptibility to kidney-related adverse responses in experimental models, supporting its relevance to nephrology and pharmacokinetic research.

OAT1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC22A6 expression without altering the underlying DNA sequence.

OAT1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC22A6 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 SLC22A6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OAT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC22A6 locus and enabling the study of OAT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OAT1 pathway restoration in tumor cells with silenced or reduced SLC22A6 expression.

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