G6PT CRISPR Activation Plasmid (h): sc-406483-ACT

Human SLC37A4 encodes glucose-6-phosphate transporter (G6PT), an endoplasmic reticulum membrane protein that imports glucose-6-phosphate into the ER lumen to support glucose-6-phosphatase activity and metabolic homeostasis. This transport step is central to glycogenolysis and gluconeogenesis, linking cytosolic carbohydrate flux to ER-localized phosphate handling and glucose production pathways. G6PT function influences intracellular energy balance and cellular responses to nutrient availability, with downstream effects on neutrophil and hepatic metabolism. Pathogenic SLC37A4 variants are associated with glycogen storage disease type Ib, making this locus relevant for mechanistic studies of metabolic dysregulation and immune-related phenotypes.

G6PT CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC37A4 expression without altering the underlying DNA sequence.

G6PT CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC37A4 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 SLC37A4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous G6PT expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC37A4 locus and enabling the study of G6PT-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of G6PT pathway restoration in tumor cells with silenced or reduced SLC37A4 expression.

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