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bioRxiv · 10.1101/2020.09.15.298174

Stable transplantation of human mitochondrial DNA by high-throughput, pressurized mitochondrial delivery

Sercel, A. J.·Patananan, A. N.·Man, T.·Wu, T.-H.·Yu, A. K.·Guyot, G. W.·Rabizadeh, S.·Niazi, K. R.·Chiou, P.-Y.·Teitell, M. A.
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Abstract

Generating mammalian cells with specific mtDNA-nDNA combinations is desirable but difficult to achieve and would be enabling for studies of mitochondria-nucleus communication and coordination in controlling cell fates and functions. We developed MitoPunch, a force-actuated mitochondrial transfer device, to deliver isolated mitochondria into numerous target mammalian cells simultaneously. MitoPunch and MitoCeption, an alternative force-based mitochondrial transfer approach, both yield stable isolated mitochondrial recipient (SIMR) cells that permanently retain exogenous mtDNA, whereas coincubation of mitochondria with cells does not yield SIMR cells. Although a typical MitoPunch or MitoCeption delivery results in dozens of immortalized SIMR clones with restored oxidative phosphorylation, only MitoPunch can produce replication-limited, non-immortal human SIMR clones. The MitoPunch device is versatile, inexpensive to assemble, and easy to use for engineering mtDNA-nDNA combinations to enable fundamental studies and potential translational applications.

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Sercel, A. J., Patananan, A. N., Man, T., Wu, T.-H., Yu, A. K., Guyot, G. W., Rabizadeh, S., Niazi, K. R., Chiou, P.-Y., Teitell, M. A.. 2020-09-15. Stable transplantation of human mitochondrial DNA by high-throughput, pressurized mitochondrial delivery. https://doi.org/10.1101/2020.09.15.298174

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