Single-cell Individual Complete mtDNA Sequencing Uncovers Hidden Mitochondrial Heterogeneity in Human and Mouse Oocytes
Ramos Mandujano, Gerardo
Izpisua Belmonte, Juan Carlos
KAUST DepartmentBioscience Program
Biological and Environmental Sciences and Engineering (BESE) Division
Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), Thuwal 23955-6900, Kingdom of Saudi Arabia.
Permanent link to this recordhttp://hdl.handle.net/10754/666791
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AbstractThe ontogeny and dynamics of mtDNA heteroplasmy remain unclear due to limitations of current mtDNA sequencing methods. We developed individual Mitochondrial Genome sequencing (iMiGseq) of full-length mtDNA for ultra-sensitive variant detection, complete haplotyping, and unbiased evaluation of heteroplasmy levels, all at the individual mtDNA molecule level. iMiGseq uncovers unappreciated levels of heteroplasmic variants in single healthy human oocytes well below the current 1% detection limit, of which numerous variants are detrimental and could contribute to late-onset mitochondrial disease and cancer. Extreme mtDNA heterogeneity among oocytes of the same mouse female, and a strong selection against deleterious mutations in human oocytes are observed. iMiGseq could comprehensively characterize and haplotype single-nucleotide and structural variants of mtDNA and their genetic linkage in NARP/Leigh syndrome patient-derived cells. Therefore, iMiGseq could not only elucidate the mitochondrial etiology of diseases, but also help diagnose and prevent mitochondrial diseases with unprecedented precision.
CitationBi, C., Wang, L., Fan, Y., Ramos-Mandujano, G., Yuan, B., Zhou, X., … Li, M. (2020). Single-cell Individual Complete mtDNA Sequencing Uncovers Hidden Mitochondrial Heterogeneity in Human and Mouse Oocytes. doi:10.1101/2020.12.28.424537
SponsorsWe thank members of the Li laboratory, Khaled Alsayegh, Samhan Alsolami for helpful discussions; Jinna Xu and Marie Krenz Y. Sicat for administrative support. We thank Chenyang Geng at the BIOPIC core facility at Peking university for technical assistance in PacBio sequencing. We thank Professor Jasmeen Merzaban’s lab and KAUST Animal Research Core Laboratory for sharing mouse strains. The research of the Li laboratory was supported byKAUST Office of Sponsored Research (OSR), under award number BAS/1/1080-01. The work was supported by a KAUST Competitive Research Grant (award number URF/1/3412-01-01) given to ML, YH and JCIB; the National Key R&D Program of China (2016YFC1000601) and the National Natural Science Funds (81571400, 81771580 and 81971381), MMAAP foundation to YY; and the National Key Research and Development Program of China (2019YFA0110804, 2018YFC1003203), and the National Natural Science Foundation of China (81871162) to YF
PublisherCold Spring Harbor Laboratory