Improved discovery of genetic interactions using CRISPRiSeq across multiple environments.

Abstract:

:Large-scale genetic interaction (GI) screens in yeast have been invaluable for our understanding of molecular systems biology and for characterizing novel gene function. Owing in part to the high costs and long experiment times required, a preponderance of GI data has been generated in a single environmental condition. However, an unknown fraction of GIs may be specific to other conditions. Here, we developed a pooled-growth CRISPRi-based sequencing assay for GIs, CRISPRiSeq, which increases throughput such that GIs can be easily assayed across multiple growth conditions. We assayed the fitness of approximately 17,000 strains encompassing approximately 7700 pairwise interactions in five conditions and found that the additional conditions increased the number of GIs detected nearly threefold over the number detected in rich media alone. In addition, we found that condition-specific GIs are prevalent and improved the power to functionally classify genes. Finally, we found new links during respiratory growth between members of the Ras nutrient-sensing pathway and both the COG complex and a gene of unknown function. Our results highlight the potential of conditional GI screens to improve our understanding of cellular genetic networks.

journal_name

Genome Res

journal_title

Genome research

authors

Jaffe M,Dziulko A,Smith JD,St Onge RP,Levy SF,Sherlock G

doi

10.1101/gr.246603.118

subject

Has Abstract

pub_date

2019-04-01 00:00:00

pages

668-681

issue

4

eissn

1088-9051

issn

1549-5469

pii

gr.246603.118

journal_volume

29

pub_type

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