DGAT1 CRISPR Activation Plasmid (h): sc-401735-ACT

Diacylglycerol O-acyltransferase 1 (DGAT1) is an endoplasmic reticulum–localized enzyme that catalyzes the terminal step of triacylglycerol synthesis by acylating diacylglycerol with fatty acyl-CoA. Through this committed reaction in neutral lipid production, DGAT1 coordinates lipid droplet biogenesis, fatty acid buffering, and cellular energy storage, intersecting with glycerolipid metabolism and ER stress-adaptation programs. DGAT1 activity influences lipoprotein assembly and intestinal lipid absorption, linking its regulation to systemic lipid handling. Altered DGAT1 expression or function has been associated with metabolic dysregulation and lipid-driven cellular stress phenotypes relevant to obesity, insulin resistance, and hepatic steatosis research models.

DGAT1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DGAT1 expression without altering the underlying DNA sequence.

DGAT1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DGAT1 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 DGAT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DGAT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DGAT1 locus and enabling the study of DGAT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DGAT1 pathway restoration in tumor cells with silenced or reduced DGAT1 expression.

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