A Systematic Mammalian Genetic Interaction Map Reveals Pathways Underlying Ricin Susceptibility

被引:271
作者
Bassik, Michael C. [1 ,2 ]
Kampmann, Martin [1 ,2 ]
Lebbink, Robert Jan [3 ,4 ]
Wang, Shuyi [1 ,2 ]
Hein, Marco Y. [5 ]
Poser, Ina [6 ]
Weibezahn, Jimena [1 ,2 ]
Horlbeck, Max A. [1 ,2 ]
Chen, Siyuan [7 ]
Mann, Matthias [5 ]
Hyman, Anthony A. [6 ]
LeProust, Emily M. [7 ]
McManus, Michael T. [3 ,4 ]
Weissman, Jonathan S. [1 ,2 ]
机构
[1] Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, Calif Inst Quantitat Biomed Res, San Francisco, CA 94122 USA
[2] Univ Calif San Francisco, Howard Hughes Med Inst, San Francisco, CA 94122 USA
[3] Univ Calif San Francisco, Dept Microbiol & Immunol, San Francisco, CA 94122 USA
[4] Univ Calif San Francisco, Ctr Diabet, San Francisco, CA 94122 USA
[5] Max Planck Inst Biochem, D-82152 Martinsried, Germany
[6] Max Planck Inst Mol Cell Biol & Genet, D-01307 Dresden, Germany
[7] Agilent Technol, Genom Solut Unit, Santa Clara, CA 95051 USA
关键词
ENDOPLASMIC-RETICULUM; TRANSLATION INITIATION; RETROGRADE TRANSPORT; SHRNA LIBRARIES; PROTEIN TOXINS; OVARIAN-CANCER; HUMAN-CELLS; E; COLI; RNAI; ORGANIZATION;
D O I
10.1016/j.cell.2013.01.030
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:909 / 922
页数:14
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