MBOAT5 CRISPR Activation Plasmid (h): sc-403597-ACT

LPCAT3 (also known as MBOAT5) encodes a membrane-bound O-acyltransferase that remodels phospholipids through Lands’ cycle, catalyzing acyl-chain transfer to lysophosphatidylcholine to help maintain membrane composition and signaling lipid availability. By shaping polyunsaturated phospholipid pools, MBOAT5 influences membrane-dependent processes including vesicular trafficking, receptor signaling, and oxidative stress responses linked to lipid peroxidation sensitivity. Altered phospholipid remodeling has been associated with metabolic and inflammatory phenotypes, making MBOAT5 a relevant node for studying lipid homeostasis and stress-adaptation programs. Functional interrogation of this pathway is commonly applied to models of liver and cardiometabolic dysfunction, immune activation, and lipid-driven cell-state transitions.

MBOAT5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LPCAT3 expression without altering the underlying DNA sequence.

MBOAT5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LPCAT3 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 LPCAT3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MBOAT5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LPCAT3 locus and enabling the study of MBOAT5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MBOAT5 pathway restoration in tumor cells with silenced or reduced LPCAT3 expression.

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