ACSL4 CRISPR Activation Plasmid (m): sc-424503-ACT

Mouse Acsl4 encodes acyl-CoA synthetase long-chain family member 4 (ACSL4), a membrane-associated enzyme that activates long-chain polyunsaturated fatty acids by converting them to acyl-CoA thioesters for incorporation into phospholipids and downstream lipid metabolic pathways. ACSL4 is a key determinant of phospholipid remodeling and redox-sensitive membrane composition, linking lipid utilization to oxidative stress responses and ferroptosis susceptibility. Through regulation of arachidonic acid and adrenic acid metabolism, ACSL4 influences inflammatory lipid signaling, mitochondrial function, and membrane dynamics. Altered ACSL4 activity and expression have been associated with lipid peroxidation-driven pathology and has been studied in contexts including neurodegeneration, metabolic dysfunction, and cancer cell stress adaptation.

ACSL4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Acsl4 expression without altering the underlying DNA sequence.

ACSL4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Acsl4 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 Acsl4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACSL4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Acsl4 locus and enabling the study of ACSL4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACSL4 pathway restoration in tumor cells with silenced or reduced Acsl4 expression.

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