Phenotypic Profiling of the Human Genome by Time-Lapse Microscopy Reveals Genes with Functions

SEMINAIRE SPECIAL Jan ELLENBERG

Date et heure du Séminaire: 3 avril à 16 h (Auditorium)

Phenotypic profiling of the human genome by time-lapse microscopy reveals genes with functions in cell division, survival or migration

Jan Ellenberg

European Molecular Biology Laboratory (EMBL)

Gene Expression Unit

Meyerhofstr. 1

D-69117 Heidelberg

Germany

Abstract:

Despite our exponentially growing knowledge about the human genome, ranging from the primary DNA sequence to its variation and evolution, we do not know all human genes required for some of the most basic functions of life including survival, division, or migration of cells. To start to fill this gap in our knowledge we developed a high throughput phenotypic screening platform combining potent gene silencing by RNA interference and time-lapse microscopy. Time-lapse imaging of living human cells allows one of the most detailed and sensitive readouts of perturbations of cellular functions and immediately reveals the physiology of the phenotype. We have therefore carried out a genome-wide phenotypic profiling of each of the ~23 000 human protein coding genes by two day live imaging of fluorescently labeled chromosomes. Phenotypes were scored quantitatively by automatic image processing of the resulting set of at least six time-lapse sequences per human gene, i.e. a total of ~190 000 movies, and analyzed by clustering of their time-resolved phenotypic signatures. This allowed us to annotate human genes with diverse biological functions including cell survival, division and migration. We were thus able to assign functions to 13% of the human genome in a single experiment. We provide in depth analysis of cell division phenotypes as part of the systematic approach to study mitosis within the Mitocheck consortium and will make the entire high content data set available as a resource to the community at

Selected recent publication:

Neumann, B., M. Held, U. Liebel, H. Erfle, P. Rogers, R. Pepperkok, and J. Ellenberg. 2006. High-throughput RNAi screening by time-lapse imaging of live human cells. Nat Methods. 3:385-90.

Lenart, P., C.P. Bacher, N. Daigle, A.R. Hand, R. Eils, M. Terasaki, and J. Ellenberg. 2005. A contractile nuclear actin network drives chromosome congression in oocytes. Nature. 436:812-8.

Rabut, G., V. Doye, and J. Ellenberg. 2004. Mapping the dynamic organization of the nuclear pore complex inside single living cells. Nat Cell Biol. 6:1114-21.

Gerlich, D., J. Beaudouin, B. Kalbfuss, N. Daigle, R. Eils, and J. Ellenberg. 2003. Global Chromosome Positions Are Transmitted through Mitosis in Mammalian Cells. Cell. 112:751-64.

Beaudouin, J., D. Gerlich, N. Daigle, R. Eils, and J. Ellenberg. 2002. Nuclear envelope breakdown proceeds by microtubule-induced tearing of the lamina. Cell. 108:83-96.