ABCD2 CRISPR Activation Plasmid (h): sc-405099-ACT

ABCD2 encodes a peroxisomal ATP-binding cassette (ABC) transporter that imports very long-chain fatty acids and related acyl-CoAs into peroxisomes for β-oxidation, supporting cellular lipid homeostasis and redox balance. In human cells, ABCD2 acts in concert with other peroxisomal transporters to regulate fatty acid catabolism, membrane lipid remodeling, and metabolic adaptation under oxidative or inflammatory stress. Altered peroxisomal lipid handling and dysregulated very long-chain fatty acid metabolism are implicated in neurodegenerative and leukodystrophy-related mechanisms, making ABCD2 a useful gene for studying compensatory pathways in peroxisome biology. ABCD2 expression and activity are also relevant to mitochondrial–peroxisomal crosstalk, including downstream effects on energy metabolism and lipid signaling.

ABCD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ABCD2 expression without altering the underlying DNA sequence.

ABCD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ABCD2 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 ABCD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ABCD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ABCD2 locus and enabling the study of ABCD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ABCD2 pathway restoration in tumor cells with silenced or reduced ABCD2 expression.

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