COX5a CRISPR Activation Plasmid (h): sc-405025-ACT

COX5A encodes cytochrome c oxidase subunit 5A (COX5a), a nuclear-encoded component of mitochondrial Complex IV that supports electron transfer to oxygen and efficient oxidative phosphorylation. By modulating cytochrome c oxidase assembly and activity, COX5a influences respiratory chain flux, mitochondrial membrane potential, and ATP production, with downstream effects on ROS signaling and metabolic adaptation. Altered Complex IV function is broadly relevant to mitochondrial bioenergetic dysfunction, which has been implicated in neurodegeneration, cardiometabolic phenotypes, and tumor cell metabolic remodeling. COX5A expression and Complex IV integrity are therefore commonly examined in studies of mitochondrial stress responses, hypoxia adaptation, and energy-dependent cell fate decisions.

COX5a CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous COX5A expression without altering the underlying DNA sequence.

COX5a CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the COX5A 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 COX5A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous COX5a expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native COX5A locus and enabling the study of COX5a-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of COX5a pathway restoration in tumor cells with silenced or reduced COX5A expression.

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