MCT3 CRISPR Activation Plasmid (h): sc-403973-ACT

SLC16A8 encodes monocarboxylate transporter 3 (MCT3), a proton-coupled transporter that mediates transmembrane flux of lactate and other short-chain monocarboxylates to support cellular redox balance and pH homeostasis. MCT3 activity contributes to metabolic coupling between cells by facilitating lactate handling and intersects with glycolytic and oxidative metabolism, influencing intracellular acidification and energy substrate availability. In the human eye, SLC16A8 is enriched in retinal pigment epithelium where it supports lactate efflux from the outer retina and helps maintain the metabolic microenvironment of photoreceptors. Genetic variation and altered expression of SLC16A8 have been associated with retinal degeneration phenotypes and age-related macular degeneration risk in human genetic studies, motivating mechanistic investigation of lactate transport in retinal health and stress responses.

MCT3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC16A8 expression without altering the underlying DNA sequence.

MCT3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC16A8 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 SLC16A8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MCT3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC16A8 locus and enabling the study of MCT3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MCT3 pathway restoration in tumor cells with silenced or reduced SLC16A8 expression.

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