GPD1 CRISPR Activation Plasmid (h): sc-403706-ACT

Human GPD1 encodes cytosolic glycerol-3-phosphate dehydrogenase 1, an NADH-dependent enzyme that interconverts dihydroxyacetone phosphate and glycerol-3-phosphate to couple carbohydrate metabolism with lipid biosynthesis. This activity supports the glycerol-3-phosphate shuttle, contributes to redox homeostasis, and supplies glycerol-3-phosphate for triglyceride and phospholipid formation. Through its role at the interface of glycolysis, glycerolipid metabolism, and cellular NADH/NAD+ balance, GPD1 helps shape metabolic flux under nutrient and hypoxic stress. Dysregulated GPD1-associated pathways have been linked in the literature to metabolic phenotypes involving hepatic lipid handling and broader energy homeostasis, making it a useful node for mechanistic studies of metabolic regulation.

GPD1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPD1 expression without altering the underlying DNA sequence.

GPD1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPD1 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 GPD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPD1 locus and enabling the study of GPD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPD1 pathway restoration in tumor cells with silenced or reduced GPD1 expression.

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