Measuring genetic interactions in human cells by RNAi and imaging

被引：15
|
作者：
Laufer, Christina [1 ,2 ]
Fischer, Bernd [3 ]
Huber, Wolfgang [3 ]
Boutros, Michael [1 ,2 ]
机构：
[1] German Canc Res Ctr, Div Signaling & Funct Genom, Heidelberg, Germany
[2] Heidelberg Univ, Dept Cell & Mol Biol, Heidelberg, Germany
[3] European Mol Biol Lab, Genome Biol Unit, D-69012 Heidelberg, Germany
来源：
NATURE PROTOCOLS | 2014年 / 9卷 / 10期
基金：
欧洲研究理事会;
关键词：
NETWORKS; PHENOTYPES;
D O I：
10.1038/nprot.2014.160
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Observation of how genetic interactions modulate phenotypes is a powerful method for dissecting their underlying molecular and functional networks. Whereas in model organisms genetic interaction studies are well established, systematic analysis of genetic interactions in human cells has remained challenging. Here we provide a detailed protocol for large-scale mapping of genetic interactions in human cells using a high-throughput phenotyping approach. Pairwise gene product depletion is induced by siRNA-mediated knockdown, and the resulting phenotypes are quantified by automated imaging and computational analysis to provide the basis for detecting genetic interactions between all pairs of genes tested. The whole workflow, depending on the size of the experiment, takes 3 or more weeks to complete.
引用
收藏
页码：2341 / 2353
页数：13
相关论文
共 50 条
  • [31] Measuring Implicit Attitudes in Human-Computer Interactions
    Kiselev, Andrey
    Abdikeev, Niyaz
    Nishida, Toyoaki
    ROUGH SETS, FUZZY SETS, DATA MINING AND GRANULAR COMPUTING, RSFDGRC 2011, 2011, 6743 : 350 - 357
  • [32] Measuring human hematopoietic stem cells
    Ando, Kiyoshi
    Yahata, Takashi
    BLOOD, 2006, 108 (02) : 771 - 772
  • [33] Downregulation of survivin by RNAi inhibits growth of human gastric carcinoma cells
    Miao, Guo-Ying
    Lu, Qi-Ming
    Zhang, Xiu-Lan
    WORLD JOURNAL OF GASTROENTEROLOGY, 2007, 13 (08) : 1170 - 1174
  • [34] RNAi in human screens
    Cannell, E
    EUROPEAN JOURNAL OF CANCER, 2004, 40 (09) : 1284 - 1284
  • [35] Forward RNAi screens in primary human hematopoietic stem/progenitor cells
    Ali, Nicole
    Karlsson, Christine
    Aspling, Marie
    Hu, Guang
    Hacohen, Nir
    Scadden, David T.
    Larsson, Jonas
    BLOOD, 2009, 113 (16) : 3690 - 3695
  • [36] Downregulation of survivin by RNAi inhibits growth of human gastric carcinoma cells
    Guo-Ying Miao
    World Journal of Gastroenterology, 2007, (08) : 1170 - 1174
  • [37] RNAi depletion of deoxycytidine and deoxyguanosine kinase in human leukemic CEM cells
    Fyrberg, Anna
    Albertioni, Freidoun
    Lotfi, Kourosh
    NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS, 2008, 27 (6-7): : 712 - 719
  • [38] Profiling essential genes in human mammary cells by multiplex RNAi screening
    Silva, Jose M.
    Marran, Krista
    Parker, Joel S.
    Silva, Javier
    Golding, Michael
    Schlabach, Michael R.
    Elledge, Stephen J.
    Hannon, Gregory J.
    Chang, Kenneth
    SCIENCE, 2008, 319 (5863) : 617 - 620
  • [39] Combinatorial interactions of genetic variants in human cardiomyopathy
    Deacon, Dekker C.
    Happe, Cassandra L.
    Chen, Chao
    Tedeschi, Neil
    Manso, Ana Maria
    Li, Ting
    Dalton, Nancy D.
    Peng, Qian
    Farah, Elie N.
    Gu, Yusu
    Tenerelli, Kevin P.
    Tran, Vivien D.
    Chen, Ju
    Peterson, Kirk L.
    Schork, Nicholasj
    Adler, Eric D.
    Engler, Adam J.
    Ross, Robert S.
    Chi, Neil C.
    NATURE BIOMEDICAL ENGINEERING, 2019, 3 (02) : 147 - 157
  • [40] Synthetic lethal genetic interactions in spindle assembly checkpoint (SAC)-deficient human cells.
    Raaijmakers, J. A.
    Van Heesbeen, R.
    Blomen, V.
    Janssen, L.
    v Diemen, F.
    Brummelkamp, T.
    Medema, R. H.
    MOLECULAR BIOLOGY OF THE CELL, 2016, 27
12345