LENG4 CRISPR Activation Plasmid (h): sc-408536-ACT

Human MBOAT7 encodes a membrane-bound O-acyltransferase that remodels phosphatidylinositol by transferring polyunsaturated acyl chains, shaping PI composition and downstream phosphoinositide signaling. Through its role in the Lands’ cycle and glycerophospholipid metabolism, MBOAT7 can influence membrane curvature, vesicular trafficking, and receptor-coupled signaling outputs that depend on PIP species. Genetic and functional studies have linked altered MBOAT7 activity to perturbations in lipid homeostasis and inflammatory signaling, with associations reported across metabolic and neurodevelopmental phenotypes. Modulating MBOAT7 expression provides a tractable approach to interrogate how PI acyl-chain remodeling impacts signaling networks, organelle function, and stress responses in human cell models.

LENG4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MBOAT7 expression without altering the underlying DNA sequence.

LENG4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MBOAT7 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 MBOAT7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LENG4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MBOAT7 locus and enabling the study of LENG4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LENG4 pathway restoration in tumor cells with silenced or reduced MBOAT7 expression.

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