ACSL4 CRISPR Activation Plasmid (h): sc-401649-ACT

Human ACSL4 (acyl‑CoA synthetase long‑chain family member 4) catalyzes the ATP-dependent activation of long-chain polyunsaturated fatty acids, generating acyl‑CoA thioesters that channel lipids into membrane remodeling and complex lipid biosynthesis. By shaping phospholipid composition, ACSL4 influences arachidonic acid metabolism, eicosanoid signaling, and lipid peroxidation dynamics that intersect with redox homeostasis and regulated cell death pathways. Altered ACSL4 activity has been associated with dysregulated lipid metabolism in cancer biology, neurodegenerative processes, and inflammatory states, making it a relevant node for studying ferroptosis sensitivity and metabolic reprogramming. Its expression and enzymatic output provide mechanistic links between lipid flux, mitochondrial function, and stress-response signaling.

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

ACSL4 CRISPR Activation Plasmid (h) 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.