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 Resjournal_title
Genome researchauthors
Jaffe M,Dziulko A,Smith JD,St Onge RP,Levy SF,Sherlock Gdoi
10.1101/gr.246603.118subject
Has Abstractpub_date
2019-04-01 00:00:00pages
668-681issue
4eissn
1088-9051issn
1549-5469pii
gr.246603.118journal_volume
29pub_type
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