SVCT1 CRISPR Activation Plasmid (h): sc-403006-ACT

SLC23A1 encodes the human sodium-dependent vitamin C transporter 1 (SVCT1), a plasma membrane symporter that mediates high-capacity uptake of L-ascorbate coupled to sodium gradients. SVCT1 activity supports cellular redox homeostasis by maintaining intracellular ascorbate pools that influence antioxidant defense, collagen hydroxylation, and dioxygenase-dependent epigenetic regulation. In epithelial tissues such as intestine and kidney, SVCT1 contributes to dietary absorption and renal reabsorption of vitamin C, shaping systemic ascorbate availability. Altered SLC23A1 function or expression has been investigated in contexts of oxidative stress susceptibility and metabolic and inflammatory phenotypes where vitamin C transport can modulate stress-response signaling pathways.

SVCT1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC23A1 expression without altering the underlying DNA sequence.

SVCT1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC23A1 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 SLC23A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SVCT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC23A1 locus and enabling the study of SVCT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SVCT1 pathway restoration in tumor cells with silenced or reduced SLC23A1 expression.

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