| 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: |
 Click here to retrieve PDF version of paper (441KB) |
