ACOX1 CRISPR Activation Plasmid (m2): sc-418948-ACT-2

Mouse Acox1 encodes acyl-CoA oxidase 1 (ACOX1), a peroxisomal flavoprotein that catalyzes the first and rate-limiting step of straight-chain fatty acyl-CoA β-oxidation, generating trans-2-enoyl-CoA and contributing to peroxisome-dependent lipid catabolism. ACOX1 activity supports cellular lipid and energy homeostasis and intersects with peroxisome biogenesis, oxidative stress regulation via H2O2 production, and metabolic signaling networks such as PPAR-driven transcriptional programs. Dysregulation of Acox1 is linked to altered very-long-chain fatty acid turnover, hepatic lipid accumulation, inflammation, and broader peroxisomal metabolism phenotypes, making it relevant to studies of fatty liver disease mechanisms and metabolic remodeling. Gene editing or genetic perturbation of mouse Acox1 is useful for dissecting peroxisomal β-oxidation pathways, lipidomics signatures, and organ- or cell-type-specific consequences of peroxisome dysfunction in biomedical research models.

ACOX1 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Acox1 expression without altering the underlying DNA sequence.

ACOX1 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Acox1 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 Acox1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACOX1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Acox1 locus and enabling the study of ACOX1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACOX1 pathway restoration in tumor cells with silenced or reduced Acox1 expression.

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