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Publication Citation: Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria

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Author(s): Joseph E. Reiner; Rani Kishore; Barbara C. Levin; Thomas Albanetti; Nicholas Boire; Ashley Knipe; Kristian Helmerson; Koren Deckman;
Title: Detection of Heteroplasmic Mitochondrial DNA in Single Mitochondria
Published: December 17, 2010
Abstract: Background Mitochondrial DNA (mtDNA) genome mutations can lead to energy and respiratory-related disorders like myoclonic epilepsy with ragged red fiber disease (MERRF), mitochondrial myopathy, encephalopathy, lactic acidosis and stroke (MELAS) syndrome, and Leber s hereditary optic neuropathy (LHON). It is not well understood what effect the distribution of mutated mtDNA throughout the mitochondrial matrix has on the development of mitochondrial-based disorders. Insight into this complex sub-cellular heterogeneity may further our understanding of the development of mitochondria-related diseases. Methodology This work describes a method for isolating individual mitochondria from single cells and performing molecular analysis on that single mitochondrion s DNA. An optical tweezer extracts a single mitochondrion from a lysed human HL-60 cell. Then a micron-sized femtopipette tip captures the mitochondrion for subsequent analysis. Multiple rounds of conventional DNA amplification and standard sequencing methods enable the detection of a heteroplasmic mixture in the mtDNA from a single mitochondrion. Significance Molecular analysis of mtDNA from the individually extracted mitochondrion demonstrates that a heteroplasmy is present in single mitochondria at various ratios consistent with the 50/50 heteroplasmy ratio found in single cells that contain multiple mitochondria.
Citation: PLoS One
Volume: 5
Issue: 12
Pages: 9 pp.
Keywords: nanobiotechnology; optical tweezers; mitochondria; DNA technologies
Research Areas: Life Sciences Research
PDF version: PDF Document Click here to retrieve PDF version of paper (451KB)