ACADVL (VLCAD) CRISPR Activation Plasmid (h): sc-403111-ACT

ACADVL encodes very long-chain acyl-CoA dehydrogenase (VLCAD), a mitochondrial flavoprotein that catalyzes the initial dehydrogenation step in β-oxidation of long-chain fatty acyl-CoAs. VLCAD activity supports cellular energy homeostasis by coupling fatty acid catabolism to electron transfer via ETF/ETFDH and downstream oxidative phosphorylation, particularly in high-energy tissues such as heart and skeletal muscle. Altered ACADVL expression or function perturbs mitochondrial fatty acid oxidation, promotes accumulation of long-chain acylcarnitines, and disrupts metabolic flexibility under fasting or increased energy demand. As a result, ACADVL is widely studied in the context of inherited fatty acid oxidation disorders and broader mitochondrial metabolic dysfunction relevant to cardiometabolic and myopathic phenotypes.

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

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

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