Glucose Transporter Glut1 CRISPR Activation Plasmid (m): sc-422998-ACT

Slc2a1 encodes glucose transporter 1 (GLUT1), a facilitative membrane transporter that mediates basal glucose uptake and helps maintain cellular energy homeostasis. In mouse tissues, GLUT1 supports glycolytic flux and pentose phosphate pathway inputs by controlling glucose availability, influencing redox balance and biosynthetic capacity. Its activity is tightly linked to metabolic adaptation under hypoxia and nutrient stress, integrating with pathways such as HIF signaling and AMPK-regulated energy sensing. Dysregulated Slc2a1/GLUT1 expression is frequently studied in contexts of altered metabolism and barrier or vascular biology, where changes in glucose transport can shift cell survival, proliferation, and inflammatory responses.

Glucose Transporter Glut1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Slc2a1 expression without altering the underlying DNA sequence.

Glucose Transporter Glut1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Slc2a1 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 Slc2a1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Glucose Transporter Glut1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Slc2a1 locus and enabling the study of Glucose Transporter Glut1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Glucose Transporter Glut1 pathway restoration in tumor cells with silenced or reduced Slc2a1 expression.

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