CNT2 CRISPR Activation Plasmid (h): sc-409209-ACT

SLC28A2 encodes concentrative nucleoside transporter 2 (CNT2), a high-affinity, sodium-dependent transporter that mediates cellular uptake of purine nucleosides such as adenosine and inosine across the plasma membrane. By controlling nucleoside salvage and intracellular nucleotide pools, CNT2 influences energy metabolism, nucleic acid synthesis, and purinergic signaling pathways that shape cellular stress responses. CNT2 activity is particularly relevant in tissues with high nucleoside flux, including intestine and kidney, and it contributes to pharmacology and toxicology contexts where nucleoside analog handling is a key variable. Dysregulated nucleoside transport has been linked to altered metabolic states and disease-associated remodeling of nucleotide homeostasis, making SLC28A2 a useful target for mechanistic studies of transport-driven pathway changes.

CNT2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC28A2 expression without altering the underlying DNA sequence.

CNT2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC28A2 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 SLC28A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CNT2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC28A2 locus and enabling the study of CNT2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CNT2 pathway restoration in tumor cells with silenced or reduced SLC28A2 expression.

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