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PLoS One. 2017 Feb 7;12(2):e0171302. doi: 10.1371/journal.pone.0171302. eCollection 2017.

Genetic interaction mapping with microfluidic-based single cell sequencing.

Author information

1
Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, United States of America.
2
Department of Bioengineering, University of California Berkeley, Berkeley, California, United States of America.
3
E.O. Lawrence Berkeley National Laboratory, Berkeley, California, United States of America.

Abstract

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.

PMID:
28170417
PMCID:
PMC5295688
DOI:
10.1371/journal.pone.0171302
[Indexed for MEDLINE]
Free PMC Article

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