cytochrome c1 CRISPR Activation Plasmid (h): sc-404958-ACT

Human CYC1 encodes cytochrome c1, a core subunit of mitochondrial respiratory chain complex III (ubiquinol–cytochrome c reductase) that mediates electron transfer from ubiquinol to cytochrome c and supports proton translocation across the inner mitochondrial membrane. Through its role in oxidative phosphorylation, cytochrome c1 contributes to ATP production, mitochondrial membrane potential maintenance, and redox homeostasis, with downstream effects on reactive oxygen species signaling and metabolic adaptation. Altered complex III function and mitochondrial bioenergetic imbalance are frequently linked to neuromuscular and metabolic phenotypes and have been observed in contexts of cancer metabolism and other disorders characterized by mitochondrial dysfunction. CYC1 expression and complex III integrity are therefore widely studied as readouts of electron transport chain regulation, mitochondrial stress responses, and bioenergetic remodeling.

cytochrome c1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYC1 expression without altering the underlying DNA sequence.

cytochrome c1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYC1 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 CYC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous cytochrome c1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYC1 locus and enabling the study of cytochrome c1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of cytochrome c1 pathway restoration in tumor cells with silenced or reduced CYC1 expression.

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