OCTN1 CRISPR Activation Plasmid (h): sc-403258-ACT

SLC22A4 encodes the human organic cation/carnitine transporter OCTN1, a polyspecific solute carrier that regulates cellular uptake and efflux of small cationic metabolites across the plasma membrane. OCTN1 contributes to nutrient handling and redox homeostasis by transporting substrates such as ergothioneine and related organic cations, influencing intracellular antioxidant capacity and metabolic adaptation. Through these transport activities, SLC22A4 intersects with membrane transport networks that shape epithelial barrier function, immune cell physiology, and xenobiotic disposition. Genetic variation and altered expression of SLC22A4 have been linked in the literature to inflammatory and autoimmune phenotypes and to changes in cellular stress responses, supporting its relevance for mechanistic studies in immunometabolism and epithelial biology.

OCTN1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC22A4 expression without altering the underlying DNA sequence.

OCTN1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC22A4 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 SLC22A4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OCTN1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC22A4 locus and enabling the study of OCTN1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OCTN1 pathway restoration in tumor cells with silenced or reduced SLC22A4 expression.

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