AGPS CRISPR Activation Plasmid (h2): sc-404604-ACT-2

Human AGPS (alkylglycerone phosphate synthase) encodes a peroxisomal enzyme that catalyzes a committed step in ether phospholipid (plasmalogen) biosynthesis by converting acyl-dihydroxyacetone phosphate to alkyl-dihydroxyacetone phosphate, thereby supporting membrane composition and lipid-mediated signaling. AGPS activity links peroxisome-dependent lipid metabolism with broader pathways regulating oxidative stress responses, organelle homeostasis, and lipid remodeling across ER and mitochondrial interfaces. Disruption of AGPS function is implicated in peroxisome biogenesis and plasmalogen-deficiency disorders, including rhizomelic chondrodysplasia punctata spectrum phenotypes, making it a relevant target for studying neurodevelopmental and skeletal pathology mechanisms. Gene editing or perturbation of AGPS is useful for generating cellular models to interrogate peroxisomal lipid flux, plasmalogen-dependent membrane dynamics, and downstream effects on cellular physiology and metabolism.

AGPS CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous AGPS expression without altering the underlying DNA sequence.

AGPS CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AGPS 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 AGPS transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AGPS expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AGPS locus and enabling the study of AGPS-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AGPS pathway restoration in tumor cells with silenced or reduced AGPS expression.

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