Arrayed adenoviral expression libraries for functional screening (original) (raw)

Nature Biotechnology volume 20, pages 1154–1157 (2002)Cite this article

A Corrigendum to this article was published on 01 February 2003

Abstract

With 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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Acknowledgements

We acknowledge the gift of mesenchymal stem cells by IsoTis (Bilthoven, The Netherlands). We thank John Collard for providing the MDCK cells. This work was supported in part by an IWT grant from the Flemish government.

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Authors and Affiliations

  1. Galapagos Genomics, Archimedesweg 4, Leiden, 2333 CN, The Netherlands
    Frits Michiels, Helmuth van Es, Kristina Dokic, Ivo Piest, Heidi Pavliska, Yvonne Rombout, Ellen Langemeijer, Libin Ma, Christel Schipper & Onno van de Stolpe
  2. Galapagos Genomics, Generaal de Wittelaan L11 A3, Mechelen, 2800, Belgium
    Luc van Rompaey, Pascal Merchiers, Bart Francken, Karen Pittois, Jan van der Schueren, Reginald Brys, Johan Vandersmissen, Filip Beirinckx, Sofie Herman, Hugo Klaassen, Evi Narinx, Annick Hagers, Wendy Laenen, Marc De Raeymaeker, Stephane Schweicher, Mia Jans, Kris van Beeck, Ing-Ren Tsang, Onno van de Stolpe & Peter Tomme

Authors

  1. Frits Michiels
  2. Helmuth van Es
  3. Luc van Rompaey
  4. Pascal Merchiers
  5. Bart Francken
  6. Karen Pittois
  7. Jan van der Schueren
  8. Reginald Brys
  9. Johan Vandersmissen
  10. Filip Beirinckx
  11. Sofie Herman
  12. Kristina Dokic
  13. Hugo Klaassen
  14. Evi Narinx
  15. Annick Hagers
  16. Wendy Laenen
  17. Ivo Piest
  18. Heidi Pavliska
  19. Yvonne Rombout
  20. Ellen Langemeijer
  21. Libin Ma
  22. Christel Schipper
  23. Marc De Raeymaeker
  24. Stephane Schweicher
  25. Mia Jans
  26. Kris van Beeck
  27. Ing-Ren Tsang
  28. Onno van de Stolpe
  29. Peter Tomme

Corresponding author

Correspondence toHelmuth van Es.

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Competing interests

All authors are employed by Galapagos Genetics, and the research described in this article was funded by Galapagos Genetics.

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Michiels, F., van Es, H., van Rompaey, L. et al. Arrayed adenoviral expression libraries for functional screening.Nat Biotechnol 20, 1154–1157 (2002). https://doi.org/10.1038/nbt746

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