ACSL1 CRISPR Activation Plasmid (m): sc-420278-ACT

Mouse Acsl1 encodes long-chain acyl-CoA synthetase 1 (ACSL1), an enzyme that activates long-chain fatty acids by converting them to acyl-CoA thioesters, thereby committing lipid substrates to β-oxidation, phospholipid remodeling, and triglyceride synthesis. ACSL1 helps coordinate cellular energy homeostasis and lipid handling across mitochondria- and ER-associated pathways, influencing processes such as lipid droplet dynamics and membrane composition. Altered ACSL1 activity and expression patterns are frequently studied in the context of metabolic inflammation, insulin resistance, hepatic steatosis, and cardiovascular-relevant lipid remodeling, where shifts in acyl-CoA pools can rewire signaling and mitochondrial function. As a result, Acsl1 is a useful node for dissecting fatty acid metabolism, nutrient sensing, and lipid-driven stress responses in mouse models and cell systems.

ACSL1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Acsl1 expression without altering the underlying DNA sequence.

ACSL1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Acsl1 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 Acsl1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACSL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Acsl1 locus and enabling the study of ACSL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACSL1 pathway restoration in tumor cells with silenced or reduced Acsl1 expression.

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