OmicsCam Enables Trimodal Profiling of Mitochondrial Genome Editing

Mitochondrial DNA (mtDNA) editing can generate cellular and animal models of mitochondrial genetic disorders and holds promise for future ex vivo and in vivo therapeutic applications. However, due to the quantitative nature of mitochondrion genetics, as more base-editing tools evolve, it is crucial to evaluate not only their efficiency and specificity on the sequence level but also the resulting molecular phenotypes. Here, we devised a novel Omics Carrier microcapsule, abbreviated as OmicsCam, that achieves homogeneous reactions within a heterogeneous carrier membrane, enabling highly efficient multistep biochemistry workflows. Incorporating magnetic beads into the carrier enables high-throughput automation. We demonstrated simultaneous trimodal assessment of mtDNA editing efficiency, postediting cellular transcriptome, and chromatin accessibility in minute cell samples containing as few as 25,000 cells. Applying OmicsCam to two TALE-DdCBE-edited human cell lines revealed that ND4 gene knockdown led to the downregulation of the mitochondrial oxidative phosphorylation pathway and changes in NF-Y transcription factor-associated histone modification pathways in the cell nucleus. Our study provides the most comprehensive analysis of mitochondrial gene editing efficiency and molecular phenotypes to date, which not only facilitates the establishment of mitochondrial genotype-molecular phenotype relationships but also helps assess the global safety of mitochondrial genome nucleases prior to clinical use.