Journal
CELL METABOLISM
Volume 23, Issue 5, Pages 921-929Publisher
CELL PRESS
DOI: 10.1016/j.cmet.2016.04.007
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Funding
- UC Discovery Biotechnology grant [178517]
- Air Force Office of Scientific Research grant [FA9550-15-1-0406]
- NIH [GM007185, GM114188, GM073981, GM061721, EB014456, CA009056, CA90571, CA009120, CA156674, CA185189, CA168585]
- NSF [CBET-1404080]
- CIRM [RB1-01397, RT3-07678]
- Prostate Cancer Foundation Challenge Award
- Broad Stem Cell Research Center Training Grant and Innovator Award
- American Cancer Society Research Scholar Award [RSG-12-257-01-TBE]
- Melanoma Research Alliance Established Investigator Award [20120279]
- National Center for Advancing Translational Sciences UCLA CTSI Grant [UL1TR000124]
- NanoCav, LLC
- NantWorks, LLC
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1404080] Funding Source: National Science Foundation
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mtDNA sequence alterations are challenging to generate but desirable for basic studies and potential correction of mtDNA diseases. Here, we report a new method for transferring isolated mitochondria into somatic mammalian cells using a photothermal nanoblade, which bypasses endocytosis and cell fusion. The nanoblade rescued the pyrimidine auxotroph phenotype and respiration of rho 0 cells that lack mtDNA. Three stable isogenic nanoblade-rescued clones grown in uridine-free medium showed distinct bioenergetics profiles. Rescue lines 1 and 3 reestablished nucleus-encoded anapleurotic and catapleurotic enzyme gene expression patterns and had metabolite profiles similar to the parent cells from which the rho 0 recipient cells were derived. By contrast, rescue line 2 retained a rho 0 cell metabolic phenotype despite growth in uridine-free selection. The known influence of metabolite levels on cellular processes, including epigenome modifications and gene expression, suggests metabolite profiling can help assess the quality and function of mtDNA-modified cells.
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