A Systematic Mammalian Genetic Interaction Map Reveals Pathways Underlying Ricin Susceptibility

作者：Bassik Michael C^*; Kampmann Martin; Lebbink Robert Jan; Wang Shuyi; Hein Marco Y; Poser Ina; Weibezahn Jimena; Horlbeck Max A; Chen Siyuan; Mann Matthias; Hyman Anthony A; LeProust Emily M; McManus Michael T; Weissman Jonathan S

来源：Cell, 2013, 152(4): 909-922.

DOI：10.1016/j.cell.2013.01.030

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.

• 出版日期2013-2-14