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Microarrays of cells expressing defined cDNAs
Naturevolume 411, pages107–110 (2001)Cite this article
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Abstract
Genome and expressed sequence tag projects are rapidly cataloguing and cloning the genes of higher organisms, including humans. An emerging challenge is to rapidly uncover the functions of genes and to identify gene products with desired properties. We have developed a microarray-driven gene expression system for the functional analysis of many gene products in parallel. Mammalian cells are cultured on a glass slide printed in defined locations with different DNAs. Cells growing on the printed areas take up the DNA, creating spots of localized transfection within a lawn of non-transfected cells. By printing sets of complementary DNAs cloned in expression vectors, we make microarrays whose features are clusters of live cells that express a defined cDNA at each location. Here we demonstrate two uses for our approach: as an alternative to protein microarrays for the identification of drug targets, and as an expression cloning system for the discovery of gene products that alter cellular physiology. By screening transfected cell microarrays expressing 192 different cDNAs, we identified proteins involved in tyrosine kinase signalling, apoptosis and cell adhesion, and with distinct subcellular distributions.
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Acknowledgements
We thank C. Wilson and T. Volkert for advice on use of the microarrayer, and K. McKernan and P. McEwan for the neurotransmitter receptor constructs. We also thank R. Young, N. Hacohen and members of the Sabatini laboratory for their support and suggestions; P. Kim and M. Gerlach for help with the manuscript; and Invitrogen for the full-length clones from the Genestorm collection. This research was supported by the G. Harold and Leila Y. Mathers Charitable Foundation, the Whitehead Institute Fellows Program and Corning. Microscopy work was conducted at the W. M. Keck Foundation Biological Imaging facility at the Whitehead Institute.
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Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, 02142, Massachusetts, USA
Junaid Ziauddin & David M. Sabatini
- Junaid Ziauddin
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- David M. Sabatini
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Correspondence toDavid M. Sabatini.
Supplementary information
Clones from the Invitrogen Genestorm collection were used to create transfected cell microarrays expressing 192 cDNAs.
Genestorm cDNAs are in pcDNA3.1/GS vector.
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Ziauddin, J., Sabatini, D. Microarrays of cells expressing defined cDNAs.Nature411, 107–110 (2001). https://doi.org/10.1038/35075114
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