NaDC-1 CRISPR Activation Plasmid (h): sc-407586-ACT

SLC13A2 encodes the sodium-dependent dicarboxylate cotransporter NaDC-1 (SLC13A2), a plasma membrane transporter that couples Na⁺ gradients to uptake of citrate, succinate, and other Krebs cycle intermediates. In polarized epithelia, NaDC-1 contributes to metabolic substrate reabsorption and intracellular citrate availability, linking membrane transport to mitochondrial metabolism, redox balance, and biosynthetic pathways. By modulating cytosolic citrate pools, SLC13A2 can influence acetyl-CoA production and downstream lipid synthesis and epigenetic acetylation processes. Altered NaDC-1 activity and dicarboxylate handling have been investigated in contexts relevant to renal tubular transport physiology, metabolic dysregulation, and disorders of organic anion homeostasis.

NaDC-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC13A2 expression without altering the underlying DNA sequence.

NaDC-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC13A2 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 SLC13A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NaDC-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC13A2 locus and enabling the study of NaDC-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NaDC-1 pathway restoration in tumor cells with silenced or reduced SLC13A2 expression.

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