OPA1 CRISPR Activation Plasmid (m): sc-428749-ACT

Mouse Opa1 encodes the mitochondrial dynamin-like GTPase OPA1, a core regulator of inner mitochondrial membrane fusion and cristae architecture. By controlling cristae junction integrity, OPA1 helps tune oxidative phosphorylation efficiency, mitochondrial DNA maintenance, and cytochrome c compartmentalization, thereby influencing intrinsic apoptosis susceptibility. OPA1 function is closely coupled to mitochondrial quality control pathways, including OMA1/YME1L-dependent proteolytic processing that balances long and short OPA1 isoforms during stress. Altered OPA1 activity is widely used as a mechanistic entry point to study mitochondrial dysfunction phenotypes relevant to neurodegeneration, optic neuropathy, and cardiometabolic stress models.

OPA1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Opa1 expression without altering the underlying DNA sequence.

OPA1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Opa1 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 Opa1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OPA1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Opa1 locus and enabling the study of OPA1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OPA1 pathway restoration in tumor cells with silenced or reduced Opa1 expression.

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