Nature Biotechnology
20, 1154 - 1157 (2002)
Published online: 30 September 2002; | doi:10.1038/nbt746
There is a Corrigenda (February 2003) associated with this Technical Report.
Arrayed adenoviral expression libraries for functional screeningFrits Michiels1, Helmuth van Es1, Luc van Rompaey2, Pascal Merchiers2, Bart Francken2, Karen Pittois2, Jan van der Schueren2, Reginald Brys2, Johan Vandersmissen2, Filip Beirinckx2, Sofie Herman2, Kristina Dokic1, Hugo Klaassen2, Evi Narinx2, Annick Hagers2, Wendy Laenen2, Ivo Piest1, Heidi Pavliska1, Yvonne Rombout1, Ellen Langemeijer1, Libin Ma1, Christel Schipper1, Marc De Raeymaeker2, Stephane Schweicher2, Mia Jans2, Kris van Beeck2, Ing-Ren Tsang2, Onno van de Stolpe1, 2
& Peter Tomme21
Galapagos Genomics, Archimedesweg 4, 2333 CN Leiden, The Netherlands. 2
Galapagos Genomics, Generaal de Wittelaan L11 A3 2800 Mechelen, Belgium.
Correspondence should be addressed to Helmuth van Es es@galapagos.beWith the publication of the sequence of the human genome, we are challenged to identify the functions of an estimated 70,000 human genes1,
2 and the much larger number of proteins encoded by these genes. Of particular interest is the identification of gene products that play a role in human disease pathways, as these proteins include potential new targets that may lead to improved therapeutic strategies. This requires the direct measurement of gene function on a genomic scale in cell-based, functional assays. We have constructed and validated an individually arrayed, replication-defective adenoviral library harboring human cDNAs, termed PhenoSelect library. The adenoviral vector guarantees efficient transduction of diverse cell types, including primary cells. The arrayed format allows screening of this library in a variety of cellular assays in search for gene(s) that, by overexpression, induce a particular disease-related phenotype. The great majority of phenotypic assays, including morphological assays, can be screened with arrayed libraries. In contrast, pooled-library approaches often rely on phenotype-based isolation or selection of single cells by employing a flow cytometer or screening for cell survival. An arrayed placental PhenoSelect library was screened in cellular assays aimed at identifying regulators of osteogenesis, metastasis, and angiogenesis. This resulted in the identification of known regulators, as well as novel sequences that encode proteins hitherto not known to play a role in these pathways. These results establish the value of the PhenoSelect platform, in combination with cellular screens, for gene function discovery.
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B and c-Jun N-terminal kinase and modulates cell growth
