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Nucleic Acids Res. 2012 Mar;40(5):2032-40. doi: 10.1093/nar/gkr949. Epub 2011 Nov 15.

Direct, genome-wide assessment of DNA mutations in single cells.

Author information

1
Department of Genetics, Albert Einstein College of Medicine, New York, NY 10461, USA.

Abstract

DNA mutations are the inevitable consequences of errors that arise during replication and repair of DNA damage. Because of their random and infrequent occurrence, quantification and characterization of DNA mutations in the genome of somatic cells has been difficult. Random, low-abundance mutations are currently inaccessible by standard high-throughput sequencing approaches because they cannot be distinguished from sequencing errors. One way to circumvent this problem and simultaneously account for the mutational heterogeneity within tissues is whole genome sequencing of a representative number of single cells. Here, we show elevated mutation levels in single cells from Drosophila melanogaster S2 and mouse embryonic fibroblast populations after treatment with the powerful mutagen N-ethyl-N-nitrosourea. This method can be applied as a direct measure of exposure to mutagenic agents and for assessing genotypic heterogeneity within tissues or cell populations.

PMID:
22086961
PMCID:
PMC3300019
DOI:
10.1093/nar/gkr949
[Indexed for MEDLINE]
Free PMC Article

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