Jul 10, 2014

Stable heteroplasmy at the single cell level is facilitated by inter-cellular exchange of mtDNA

BioRxiv : the Preprint Server for Biology
Anitha D JayaprakashRavi Sachidanandam

Abstract

Eukaryotic cells carry two genomes, nuclear (nDNA) and mitochondrial (mtDNA), which are ostensibly decoupled in their replication, segregation and inheritance. It is increasingly appreciated that heteroplasmy, the occurrence of multiple mtDNA haplotypes in a cell, plays an important biological role, but its features are not well understood. Accurately determining the diversity of mtDNA has been difficult, due to the relatively small amount of mtDNA in each cell (< 1% of the total DNA), the intercellular variability of mtDNA content and mtDNA pseudogenes (Numts) in nDNA. To understand the nature of heteroplasmy, we developed Mseek, a novel technique to purify and sequence mtDNA. Mseek yields high purity (> 90%) mtDNA and its ability to detect rare variants is limited only by sequencing depth, providing unprecedented sensitivity and specificity. Using Mseek, we confirmed the ubiquity of heteroplasmy by analyzing mtDNA from a diverse set of cell lines and human samples. Applying Mseek to colonies derived from single cells, we find heteroplasmy is stably maintained in individual daughter cells over multiple cell divisions. We hypothesized that the stability of heteroplasmy could be facilitated by inter-cellular exchange of mtDNA. W...Continue Reading

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Mentioned in this Paper

Genome
DNA, Mitochondrial
Coculture Techniques
Cell Division
Extracellular Space
Virus Replication
Nucleic Acid Sequencing
Genetic Inheritance
Sequencing
Mitochondria

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