G3PP/PGP CRISPR Activation Plasmid (h): sc-402332-ACT

Human PGP, also known as G3PP (glycerol-3-phosphate phosphatase), encodes a cytosolic phosphatase that hydrolyzes glycerol-3-phosphate to glycerol, linking glycerolipid metabolism with glycolytic flux and redox homeostasis. By modulating glycerol-3-phosphate availability, G3PP/PGP influences triglyceride synthesis, the glycerol phosphate shuttle, and mitochondrial NADH reoxidation, thereby shaping cellular responses to nutrient excess and oxidative stress. This metabolic control is relevant to studies of insulin secretion and sensitivity, lipotoxicity, and energy balance in liver, adipose tissue, skeletal muscle, and pancreatic beta cells. Dysregulated glycerolipid handling and redox pressure connected to G3PP/PGP activity are frequently examined in the context of metabolic syndrome, obesity-associated inflammation, and diabetes-related cellular dysfunction.

G3PP/PGP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PGP expression without altering the underlying DNA sequence.

G3PP/PGP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PGP 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 PGP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous G3PP/PGP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PGP locus and enabling the study of G3PP/PGP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of G3PP/PGP pathway restoration in tumor cells with silenced or reduced PGP expression.

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