OCT1 CRISPR Activation Plasmid (h): sc-402232-ACT

SLC22A1 encodes organic cation transporter 1 (OCT1), a polyspecific solute carrier that mediates sodium-independent uptake of endogenous amines and diverse xenobiotic organic cations across the plasma membrane. OCT1 activity shapes cellular disposition of charged metabolites and drugs, influencing hepatic handling of circulating compounds and contributing to interindividual variability in transport-dependent pharmacokinetics. By controlling intracellular availability of cationic substrates, OCT1 interfaces with detoxification and metabolic homeostasis pathways, including processes that modulate oxidative stress and mitochondrial function. Altered SLC22A1 expression or genetic variation has been associated with differences in drug response and has been investigated in liver-related disorders and cancer biology as a determinant of transporter-mediated uptake.

OCT1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC22A1 expression without altering the underlying DNA sequence.

OCT1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC22A1 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 SLC22A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OCT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC22A1 locus and enabling the study of OCT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OCT1 pathway restoration in tumor cells with silenced or reduced SLC22A1 expression.

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