A systematic mammalian genetic interaction map reveals pathways underlying ricin susceptibility
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A systematic mammalian genetic interaction map reveals pathways underlying ricin susceptibility. / 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.
In: Cell, Vol. 152, No. 4, 14.02.2013, p. 909-22.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - A systematic mammalian genetic interaction map reveals pathways underlying ricin susceptibility
AU - Bassik, Michael C
AU - Kampmann, Martin
AU - Lebbink, Robert Jan
AU - Wang, Shuyi
AU - Hein, Marco Y
AU - Poser, Ina
AU - Weibezahn, Jimena
AU - Horlbeck, Max A
AU - Chen, Siyuan
AU - Mann, Matthias
AU - Hyman, Anthony A
AU - Leproust, Emily M
AU - McManus, Michael T
AU - Weissman, Jonathan S
N1 - Copyright © 2013 Elsevier Inc. All rights reserved.
PY - 2013/2/14
Y1 - 2013/2/14
N2 - 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.
AB - 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.
KW - Biological Transport
KW - Carrier Proteins
KW - Cell Line, Tumor
KW - Coat Protein Complex I
KW - Endoplasmic Reticulum
KW - Epistasis, Genetic
KW - Heptanoic Acids
KW - Humans
KW - Membrane Proteins
KW - Proto-Oncogene Proteins
KW - Pyrroles
KW - RNA, Small Interfering
KW - Ribosomal Proteins
KW - Ricin
KW - Vesicular Transport Proteins
U2 - 10.1016/j.cell.2013.01.030
DO - 10.1016/j.cell.2013.01.030
M3 - Journal article
C2 - 23394947
VL - 152
SP - 909
EP - 922
JO - Cell
JF - Cell
SN - 0092-8674
IS - 4
ER -
ID: 88589714