MGAT2 CRISPR Activation Plasmid (h): sc-406205-ACT

Human MOGAT2 encodes MGAT2 (monoacylglycerol O-acyltransferase 2), an endoplasmic reticulum membrane enzyme that catalyzes conversion of monoacylglycerol and acyl-CoA to diacylglycerol, a key intermediate in triacylglycerol synthesis. This activity links dietary lipid assimilation and cellular lipid storage to broader glycerolipid and energy homeostasis pathways, influencing lipid droplet formation and membrane lipid composition. MGAT2-dependent flux can modulate downstream signaling through diacylglycerol-derived lipid mediators and intersects with metabolic programs that shape adipose and hepatic lipid handling. Altered regulation of MOGAT2/MGAT2 has been associated with metabolic phenotypes involving dyslipidemia, insulin sensitivity, and fatty liver–related processes, making it relevant for mechanistic studies of lipid-driven pathophysiology.

MGAT2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MOGAT2 expression without altering the underlying DNA sequence.

MGAT2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MOGAT2 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 MOGAT2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MGAT2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MOGAT2 locus and enabling the study of MGAT2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MGAT2 pathway restoration in tumor cells with silenced or reduced MOGAT2 expression.

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