A systematic mammalian genetic interaction map reveals pathways underlying ricin susceptibility
Research output: Contribution to journal › Journal article › Research › peer-review
Genetic interaction (GI) maps, comprising pairwise measures of how strongly the function of one gene depends on the presence of a second, have enabled the systematic exploration of gene function in microorganisms. Here, we present a two-stage strategy to construct high-density GI maps in mammalian cells. First, we use ultracomplex pooled shRNA libraries (25 shRNAs/gene) to identify high-confidence hit genes for a given phenotype and effective shRNAs. We then construct double-shRNA libraries from these to systematically measure GIs between hits. A GI map focused on ricin susceptibility broadly recapitulates known pathways and provides many unexpected insights. These include a noncanonical role for COPI, a previously uncharacterized protein complex affecting toxin clearance, a specialized role for the ribosomal protein RPS25, and functionally distinct mammalian TRAPP complexes. The ability to rapidly generate mammalian GI maps provides a potentially transformative tool for defining gene function and designing combination therapies based on synergistic pairs.
Original language | English |
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Journal | Cell |
Volume | 152 |
Issue number | 4 |
Pages (from-to) | 909-22 |
Number of pages | 14 |
ISSN | 0092-8674 |
DOIs | |
Publication status | Published - 14 Feb 2013 |
Externally published | Yes |
- Biological Transport, Carrier Proteins, Cell Line, Tumor, Coat Protein Complex I, Endoplasmic Reticulum, Epistasis, Genetic, Heptanoic Acids, Humans, Membrane Proteins, Proto-Oncogene Proteins, Pyrroles, RNA, Small Interfering, Ribosomal Proteins, Ricin, Vesicular Transport Proteins
Research areas
ID: 88589714