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  Lineage tracking for probing heritable phenotypes at single-cell resolution

Cottinet, D., Condamine, F., Bremond, N., Griffiths, A., Rainey, P., De Visser, J., et al. (2016). Lineage tracking for probing heritable phenotypes at single-cell resolution. PLoS ONE, 11(4). doi:10.1371/journal.pone.0152395.

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Item Permalink: https://hdl.handle.net/11858/00-001M-0000-002C-3AEF-1 Version Permalink: https://hdl.handle.net/11858/00-001M-0000-002C-3AF1-A
Genre: Journal Article

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Cottinet, D.1, Author
Condamine, F., Author
Bremond, N., Author
Griffiths, A.D., Author
Rainey, P.B., Author
De Visser, J.A.G.M., Author
Baudry, J., Author
Bibette, J., Author
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1External Organizations, ou_persistent22              

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Free keywords: DNA sequence; Escherichia coli; microbiology; mutation; phenotype; population biology; starvation
 Abstract: Determining the phenotype and genotype of single cells is central to understand microbial evolution. DNA sequencing technologies allow the detection of mutants at high resolution, but similar approaches for phenotypic analyses are still lacking. We show that a drop-based millifluidic system enables the detection of heritable phenotypic changes in evolving bacterial populations. At time intervals, cells were sampled and individually compartmentalized in 100 nL drops. Growth through 15 generations was monitored using a fluorescent protein reporter. Amplification of heritable changes-via growth-over multiple generations yields phenotypically distinct clusters reflecting variation relevant for evolution. To demonstrate the utility of this approach, we follow the evolution of Escherichia coli populations during 30 days of starvation. Phenotypic diversity was observed to rapidly increase upon starvation with the emergence of heritable phenotypes. Mutations corresponding to each phenotypic class were identified by DNA sequencing. This scalable lineage-tracking technology opens the door to large-scale phenotyping methods with special utility for microbiology and microbial population biology. © 2016 Cottinet et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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 Dates: 2016
 Publication Status: Issued
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 Identifiers: DOI: 10.1371/journal.pone.0152395
BibTex Citekey: Cottinet2016
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Title: PLoS ONE
Source Genre: Journal
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Publ. Info: Public Library of Science
Pages: - Volume / Issue: 11 (4) Sequence Number: - Start / End Page: - Identifier: -