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Microarrays of lentiviruses for gene function screens in immortalized and primary cells

Nature Methods volume 3, pages 117–122 (2006)Cite this article

Abstract

Here we describe lentivirus-infected cell microarrays for the high-throughput screening of gene function in mammalian cells. To create these arrays, we cultured mammalian cells on glass slides 'printed' with lentiviruses pseudotyped as vesicular stomatitis virus glycoprotein, which encode short hairpin RNA or cDNA. Cells that land on the printed 'features' become infected with lentivirus, creating a living array of stably transduced cell clusters within a monolayer of uninfected cells. The small size of the features of the microarrays (300 μm in diameter) allows high-density spotting of lentivirus, permitting thousands of distinct parallel infections on a single glass slide. Because lentiviruses have a wide cellular tropism, including primary cells, lentivirus-infected cell microarrays can be used as a platform for high-throughput screening in a variety of cell types.

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Figure 1: LICMs made with two different types of lentiviruses.
Figure 2: LICMs are compatible with a variety of transformed and primary cell lines.
Figure 3: LICMs are compatible with the detection of complex phenotypes.
Figure 4: Complex cellular phenotypes can be detected by automated image analysis of LICM features.

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Acknowledgements

We thank S.A. Stewart (Washington University; St. Louis, Missouri) for the gift of LKO.1 puro; C. Dillon (Massachusetts Institute of Technology, Cambridge, Massachusetts) for the gift of Lentilox 3.7-CMV-RFP; and N. Hacohen (Massachusetts General Hospital; Boston, Massachusetts) for the gift of LKO.3 Thy-1.1 and primary mouse dendritic cells. A.E.C. is a Novartis fellow of the Life Sciences Research Foundation; S.M.A. is a Howard Hughes Medical Institute predoctoral fellow. This work was supported by funds from the Whitehead Institute and Pew Charitable Trusts.

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Authors and Affiliations

  1. Whitehead Institute for Biomedical Research and Massachusetts Institute of Technology Department of Biology, Nine Cambridge Center, Cambridge, 02142, Massachusetts, USA

    Steve N Bailey, Siraj M Ali, Anne E Carpenter, Caitlin O Higgins & David M Sabatini

  2. Broad Institute, 320 Charles Street, Cambridge, 02141, Massachusetts, USA

    Steve N Bailey, Siraj M Ali, Anne E Carpenter, Caitlin O Higgins & David M Sabatini

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  1. Steve N Bailey
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Correspondence to David M Sabatini.

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Bailey, S., Ali, S., Carpenter, A. et al. Microarrays of lentiviruses for gene function screens in immortalized and primary cells. Nat Methods 3, 117–122 (2006). https://doi.org/10.1038/nmeth848

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