SLC5A8 CRISPR Activation Plasmid (h): sc-403705-ACT

SLC5A8 (also known as SMCT1) encodes a sodium-coupled monocarboxylate transporter that mediates uptake of short-chain fatty acids and other monocarboxylates such as lactate, pyruvate, and nicotinate across the plasma membrane. By regulating intracellular availability of these metabolites, SLC5A8 links membrane transport to cellular energy metabolism and epigenetic control, including pathways influenced by histone deacetylase–inhibitory short-chain fatty acids. Its expression is frequently altered in cancer biology and epithelial cell models, where transporter activity can affect metabolic rewiring, differentiation programs, and stress responses. SLC5A8 is also relevant to studies of nutrient sensing and barrier tissue physiology, given its role in handling luminal monocarboxylates.

SLC5A8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC5A8 expression without altering the underlying DNA sequence.

SLC5A8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC5A8 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 SLC5A8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SLC5A8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC5A8 locus and enabling the study of SLC5A8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SLC5A8 pathway restoration in tumor cells with silenced or reduced SLC5A8 expression.

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