期刊
ELIFE
卷 10, 期 -, 页码 -出版社
ELIFE SCIENCES PUBLICATIONS LTD
DOI: 10.7554/eLife.63102
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资金
- National Institutes of Health [T32CA009120, T32GM007185, T32GM008042, R01GM114188, R01GM073981, R01CA185189, R21CA227480, P30CA016042]
- American Heart Association [18POST34080342]
- National Science Foundation [CBET 1404080]
- Air Force Office of Scientific Research [FA9550-15-1-0406]
- CIRM [RT3-07678]
The MitoPunch device allows for the delivery of isolated mitochondria into mammalian cells, creating specific mtDNA-nDNA combinations. This technology yields stable isolated mitochondrial recipient (SIMR) cells, enabling studies on mitochondrial-nuclear communication and coordination.
Generating mammalian cells with specific mitochondrial DNA (mtDNA)-nuclear DNA (nDNA) combinations is desirable but difficult to achieve and would be enabling for studies of mitochondrial-nuclear communication and coordination in controlling cell fates and functions. We developed 'MitoPunch', a pressure-driven mitochondrial transfer device, to deliver isolated mitochondria into numerous target mammalian cells simultaneously. MitoPunch and MitoCeption, a previously described 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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