ECH1 CRISPR Activation Plasmid (h): sc-406076-ACT

ECH1 (enoyl-CoA hydratase 1) is a mitochondrial enzyme that catalyzes a hydration step in the β-oxidation of fatty acids and participates in the catabolism of branched-chain and unsaturated acyl-CoA intermediates. By supporting mitochondrial lipid utilization and acetyl-CoA supply, ECH1 contributes to cellular energy homeostasis and links to broader metabolic programs that intersect with redox balance and mitochondrial function. Altered fatty acid oxidation and mitochondrial metabolic rewiring are recurrent features of metabolic disorders and cancer biology, making ECH1 a useful node for studying how lipid flux influences cell growth, stress adaptation, and metabolic signaling. ECH1 is also relevant for investigating peroxisome–mitochondria crosstalk and the handling of potentially toxic fatty acid intermediates.

ECH1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ECH1 expression without altering the underlying DNA sequence.

ECH1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ECH1 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 ECH1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ECH1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ECH1 locus and enabling the study of ECH1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ECH1 pathway restoration in tumor cells with silenced or reduced ECH1 expression.

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