Jan Ellenberg及其同事利用RNA干涉來將一個(gè)人類細(xì)胞系中大約21,000個(gè)蛋白編碼基因中的每個(gè)基因沉默,，然后利用對(duì)活的、正在分裂的細(xì)胞進(jìn)行高通過量時(shí)間延遲成像的方法來記錄結(jié)果,。
通過對(duì)每個(gè)基因至少6個(gè)時(shí)長為兩天的影片進(jìn)行計(jì)算機(jī)圖像處理,，來對(duì)表現(xiàn)型進(jìn)行量化打分。數(shù)百個(gè)基因被發(fā)現(xiàn)在有絲分裂和包括細(xì)胞存活及遷移在內(nèi)的其他細(xì)胞過程中發(fā)揮功能,。
該研究的整個(gè)數(shù)據(jù)集作為一個(gè)公共功能基因組資源發(fā)布在ｗｗｗ.ｍｉｔｏｃｈｅｃｋ.ｏｒｇ網(wǎng)站上,。(生物谷Bioon.com)
生物谷推薦原文出處：
Nature doi:10.1038/nature08869
Phenotypic profiling of the human genome by time-lapse microscopy reveals cell division genes
Beate Neumann1,12, Thomas Walter1,12, Jean-Karim Hériché5,13, Jutta Bulkescher1, Holger Erfle1,3,13, Christian Conrad1,3, Phill Rogers1,13, Ina Poser6, Michael Held1,13, Urban Liebel1,13, Cihan Cetin3, Frank Sieckmann8, Gregoire Pau9, Rolf Kabbe10, Annelie Wünsche2, Venkata Satagopam4, Michael H. A. Schmitz7, Catherine Chapuis3, Daniel W. Gerlich7, Reinhard Schneider4, Roland Eils10, Wolfgang Huber9, Jan-Michael Peters11, Anthony A. Hyman6, Richard Durbin5, Rainer Pepperkok3 & Jan Ellenberg2
MitoCheck Project Group,
Gene Expression and,
Cell Biology/Biophysics Units, Structural and,
Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, D-69117 Heidelberg, Germany
Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1HH, UK
Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany
Institute of Biochemistry, Swiss Federal Institute of Technology Zurich (ETHZ), Schafmattstrasse 18, CH-8093 Zurich, Switzerland
Leica Microsystems CMS GmbH, Am Friedensplatz 3, D-68165 Mannheim, Germany
European Bioinformatics Institute, European Molecular Biology Laboratory, Cambridge CB10 1SD, UK
Division of Theoretical Bioinformatics, German Cancer Research Center, Im Neuenheimer Feld 267, D-69120 Heidelberg, Germany
Institute for Molecular Pathology, Dr Bohr Gasse 7, A-1030 Vienna, Austria
These authors contributed equally to this work.
Present addresses: MitoCheck Project Group, European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, D-69117 Heidelberg, Germany (J.-K.H.); BIOQUANT Centre University Heidelberg, INF 267, D-69120 Heidelberg, Germany (H.E.); 3-V Biosciences GmbH, Wagistrasse 27, 8952 Schlieren, Switzerland (P.R.); Institute of Biochemistry, Swiss Federal Institute of Technology Zurich (ETHZ), Schafmattstrasse 18, CH-8093 Zurich, Switzerland (M.H.); Karlsruhe Institute of Technology KIT, Herrmann-von-Helmholtz Platz 1, D-76344 Eggenstein-Leopoldshafen, Germany (U.L.).
Despite our rapidly growing knowledge about the human genome, we do not know all of the genes required for some of the most basic functions of life. To start to fill this gap we developed a high-throughput phenotypic screening platform combining potent gene silencing by RNA interference, time-lapse microscopy and computational image processing. We carried out a genome-wide phenotypic profiling of each of the ~21,000 human protein-coding genes by two-day live imaging of fluorescently labelled chromosomes. Phenotypes were scored quantitatively by computational image processing, which allowed us to identify hundreds of human genes involved in diverse biological functions including cell division, migration and survival. As part of the Mitocheck consortium, this study provides an in-depth analysis of cell division phenotypes and makes the entire high-content data set available as a resource to the community.
