OATP8 CRISPR Activation Plasmid (h): sc-402371-ACT

SLCO1B3 encodes the human organic anion transporting polypeptide OATP8 (OATP1B3), a membrane influx transporter that mediates sodium-independent uptake of diverse endogenous and xenobiotic organic anions. It is highly relevant to hepatic and epithelial transport processes that shape cellular exposure to bile acids, bilirubin conjugates, steroid hormone metabolites, and numerous drug-like compounds, thereby influencing detoxification and metabolic clearance pathways. Altered SLCO1B3 expression or mislocalization can remodel intracellular substrate availability and downstream signaling linked to xenobiotic response, oxidative stress handling, and metabolic homeostasis. Aberrant OATP1B3 activity and ectopic expression patterns have been reported in multiple cancers and liver-associated disorders, making it a useful target for mechanistic studies of transporter-regulated phenotypes and pharmacogenomic variability.

OATP8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLCO1B3 expression without altering the underlying DNA sequence.

OATP8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLCO1B3 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 SLCO1B3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OATP8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLCO1B3 locus and enabling the study of OATP8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OATP8 pathway restoration in tumor cells with silenced or reduced SLCO1B3 expression.

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