Glucose Transporter Glut4 CRISPR Activation Plasmid (h): sc-400158-ACT

SLC2A4 encodes glucose transporter Glut4, an insulin-responsive facilitative glucose transporter that drives stimulus-dependent glucose uptake in adipocytes and skeletal muscle. Upon insulin signaling through the PI3K–AKT axis, Glut4 translocates from intracellular storage vesicles to the plasma membrane, coupling receptor activation to cellular glucose utilization and glycogen synthesis. Glut4 trafficking is also modulated by AMPK-dependent metabolic stress pathways and vesicle fusion machinery, integrating nutrient availability with energy homeostasis. Dysregulated SLC2A4 expression or impaired Glut4 translocation is implicated in insulin resistance and metabolic disease phenotypes, making it a key node for studying glucose handling in human cell models.

Glucose Transporter Glut4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLC2A4 expression without altering the underlying DNA sequence.

Glucose Transporter Glut4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLC2A4 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 SLC2A4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Glucose Transporter Glut4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLC2A4 locus and enabling the study of Glucose Transporter Glut4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Glucose Transporter Glut4 pathway restoration in tumor cells with silenced or reduced SLC2A4 expression.

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