LPAAT-β CRISPR Activation Plasmid (h): sc-405045-ACT

AGPAT2 encodes lysophosphatidic acid acyltransferase beta (LPAAT-β), an endoplasmic reticulum–associated enzyme that catalyzes the acylation of lysophosphatidic acid to form phosphatidic acid, a key intermediate in glycerophospholipid and triacylglycerol biosynthesis. By controlling phosphatidic acid availability, LPAAT-β influences membrane biogenesis, lipid droplet formation, and lipid-mediated signaling processes that intersect with metabolic stress and organelle homeostasis. AGPAT2 activity is tightly linked to adipocyte lipid storage and systemic energy balance, and loss-of-function variation is associated with congenital generalized lipodystrophy and severe metabolic dysregulation. These features make AGPAT2 a relevant target for studying lipid remodeling, adipogenesis, and disease-associated perturbations in phospholipid metabolism.

LPAAT-β CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AGPAT2 expression without altering the underlying DNA sequence.

LPAAT-β CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AGPAT2 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 AGPAT2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LPAAT-β expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AGPAT2 locus and enabling the study of LPAAT-β-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LPAAT-β pathway restoration in tumor cells with silenced or reduced AGPAT2 expression.

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