Oma1 CRISPR Activation Plasmid (h): sc-402953-ACT

Human OMA1 encodes the inner mitochondrial membrane metalloprotease Oma1, a stress-responsive regulator of mitochondrial quality control. Oma1 is activated by mitochondrial depolarization and proteotoxic stress to process key substrates such as OPA1, thereby influencing mitochondrial fusion–fission balance, cristae architecture, and oxidative phosphorylation capacity. Through its coupling to mitochondrial unfolded protein responses, mitophagy initiation, and apoptotic signaling, Oma1 integrates bioenergetic stress with organelle remodeling. Dysregulation of OMA1–OPA1 axis has been associated with altered mitochondrial dynamics and has been studied in contexts including neurodegeneration, cardiometabolic dysfunction, and cancer cell stress adaptation.

Oma1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous OMA1 expression without altering the underlying DNA sequence.

Oma1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the OMA1 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 OMA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Oma1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native OMA1 locus and enabling the study of Oma1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Oma1 pathway restoration in tumor cells with silenced or reduced OMA1 expression.

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