Genetic interaction mapping with microfluidic-based single cell sequencing.

Genetic interaction mapping is useful for understanding the molecular basis of cellular decision making, but elucidating interactions genome-wide is challenging due to the massive number of gene combinations that must be tested. Here, we demonstrate a simple approach to thoroughly map genetic intera...

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Main Authors: John R Haliburton, Wenjun Shao, Adam Deutschbauer, Adam Arkin, Adam R Abate
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2017-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0171302&type=printable
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author John R Haliburton
Wenjun Shao
Adam Deutschbauer
Adam Arkin
Adam R Abate
author_facet John R Haliburton
Wenjun Shao
Adam Deutschbauer
Adam Arkin
Adam R Abate
author_sort John R Haliburton
collection DOAJ
description Genetic interaction mapping is useful for understanding the molecular basis of cellular decision making, but elucidating interactions genome-wide is challenging due to the massive number of gene combinations that must be tested. Here, we demonstrate a simple approach to thoroughly map genetic interactions in bacteria using microfluidic-based single cell sequencing. Using single cell PCR in droplets, we link distinct genetic information into single DNA sequences that can be decoded by next generation sequencing. Our approach is scalable and theoretically enables the pooling of entire interaction libraries to interrogate multiple pairwise genetic interactions in a single culture. The speed, ease, and low-cost of our approach makes genetic interaction mapping viable for routine characterization, allowing the interaction network to be used as a universal read out for a variety of biology experiments, and for the elucidation of interaction networks in non-model organisms.
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institution DOAJ
issn 1932-6203
language English
publishDate 2017-01-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS ONE
spelling doaj-art-c1a2ca4f240a4bc9be8c0d02397943122025-08-20T03:12:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01122e017130210.1371/journal.pone.0171302Genetic interaction mapping with microfluidic-based single cell sequencing.John R HaliburtonWenjun ShaoAdam DeutschbauerAdam ArkinAdam R AbateGenetic interaction mapping is useful for understanding the molecular basis of cellular decision making, but elucidating interactions genome-wide is challenging due to the massive number of gene combinations that must be tested. Here, we demonstrate a simple approach to thoroughly map genetic interactions in bacteria using microfluidic-based single cell sequencing. Using single cell PCR in droplets, we link distinct genetic information into single DNA sequences that can be decoded by next generation sequencing. Our approach is scalable and theoretically enables the pooling of entire interaction libraries to interrogate multiple pairwise genetic interactions in a single culture. The speed, ease, and low-cost of our approach makes genetic interaction mapping viable for routine characterization, allowing the interaction network to be used as a universal read out for a variety of biology experiments, and for the elucidation of interaction networks in non-model organisms.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0171302&type=printable
spellingShingle John R Haliburton
Wenjun Shao
Adam Deutschbauer
Adam Arkin
Adam R Abate
Genetic interaction mapping with microfluidic-based single cell sequencing.
PLoS ONE
title Genetic interaction mapping with microfluidic-based single cell sequencing.
title_full Genetic interaction mapping with microfluidic-based single cell sequencing.
title_fullStr Genetic interaction mapping with microfluidic-based single cell sequencing.
title_full_unstemmed Genetic interaction mapping with microfluidic-based single cell sequencing.
title_short Genetic interaction mapping with microfluidic-based single cell sequencing.
title_sort genetic interaction mapping with microfluidic based single cell sequencing
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0171302&type=printable
work_keys_str_mv AT johnrhaliburton geneticinteractionmappingwithmicrofluidicbasedsinglecellsequencing
AT wenjunshao geneticinteractionmappingwithmicrofluidicbasedsinglecellsequencing
AT adamdeutschbauer geneticinteractionmappingwithmicrofluidicbasedsinglecellsequencing
AT adamarkin geneticinteractionmappingwithmicrofluidicbasedsinglecellsequencing
AT adamrabate geneticinteractionmappingwithmicrofluidicbasedsinglecellsequencing