Reports

Genome-Wide RNAi Screen for Host Factors Required for Intracellular Bacterial Infection

Hervé Agaisse,^1,2* Laura S. Burrack,^1* Jennifer A. Philips,² Eric J. Rubin,³ Norbert Perrimon,² Darren E. Higgins¹

Most studies of host-pathogen interactions have focused on pathogen-specific virulence determinants. Here, we report a genome-wide RNA interference screen to identify host factors required for intracellular bacterial pathogenesis. Using Drosophila cells and the cytosolic pathogen Listeria monocytogenes, we identified 305 double-stranded RNAs targeting a wide range of cellular functions that altered L. monocytogenes infection. Comparison to a similar screen with Mycobacterium fortuitum, a vacuolar pathogen, identified host factors that may play a general role in intracellular pathogenesis and factors that specifically affect access to the cytosol by L. monocytogenes.

¹ Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.
² Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
³ Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, MA 02115, USA.

^* These authors contributed equally to this work.

Present address: Section of Microbial Pathogenesis, Yale University School of Medicine, New Haven, CT 06519, USA.

To whom correspondence should be addressed. E-mail: herve.agaisse{at}yale.edu (H.A.); dhiggins{at}hms.harvard.edu (D.E.H.)

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A. Schwegmann and F. Brombacher (2008)
Science Signaling 1, re8
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Development 135, 1735-1743
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Use of Drosophila S2 Cells as a Model for Studying Ehrlichia chaffeensis Infections.

A. Luce-Fedrow, T. Von Ohlen, D. Boyle, R. R. Ganta, and S. K. Chapes (2008)
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J. S. Ayres, N. Freitag, and D. S. Schneider (2008)
Genetics 178, 1807-1815
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J. A. Philips, M. C. Porto, H. Wang, E. J. Rubin, and N. Perrimon (2008)
PNAS 105, 3070-3075
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I. C. Koo, Y. M. Ohol, P. Wu, J. H. Morisaki, J. S. Cox, and E. J. Brown (2008)
PNAS 105, 710-715
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S. K. Sharma and M. Nirenberg (2007)
PNAS 104, 12925-12930
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Cancer genomics: integrating form and function.

S. Y. Kim and W. C. Hahn (2007)
Carcinogenesis 28, 1387-1392
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Knockdown of Actin and Caspase Gene Expression by RNA Interference in the Symbiotic Anemone Aiptasia pallida.

S. R. Dunn, W. S. Phillips, D. R. Green, and V. M. Weis (2007)
Biol. Bull. 212, 250-258
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Downregulation of CD4 by Human Immunodeficiency Virus Type 1 Nef Is Dependent on Clathrin and Involves Direct Interaction of Nef with the AP2 Clathrin Adaptor.

R. Chaudhuri, O. W. Lindwasser, W. J. Smith, J. H. Hurley, and J. S. Bonifacino (2007)
J. Virol. 81, 3877-3890
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PhenomicDB: a new cross-species genotype/phenotype resource.

P. Groth, N. Pavlova, I. Kalev, S. Tonov, G. Georgiev, H.-D. Pohlenz, and B. Weiss (2007)
Nucleic Acids Res. 35, D696-D699
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Applications of High-Throughput RNA Interference Screens to Problems in Cell and Developmental Biology.

N. Perrimon and B. Mathey-Prevot (2007)
Genetics 175, 7-16
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Genome-wide functional analysis of human cell-cycle regulators.

M. Mukherji, R. Bell, L. Supekova, Y. Wang, A. P. Orth, S. Batalov, L. Miraglia, D. Huesken, J. Lange, C. Martin, et al. (2006)
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V. Moos, D. Kunkel, T. Marth, G. E. Feurle, B. LaScola, R. Ignatius, M. Zeitz, and T. Schneider (2006)
J. Immunol. 177, 2015-2022
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Genome-wide RNAi screen of Ca2+ influx identifies genes that regulate Ca2+ release-activated Ca2+ channel activity.

S. L. Zhang, A. V. Yeromin, X. H.-F. Zhang, Y. Yu, O. Safrina, A. Penna, J. Roos, K. A. Stauderman, and M. D. Cahalan (2006)
PNAS 103, 9357-9362
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Cellular phenotyping by RNAi.

F. Fuchs and M. Boutros (2006)
Brief Funct Genomic Proteomic 5, 52-56
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Drosophila RNAi Screen Reveals CD36 Family Member Required for Mycobacterial Infection.

J. A. Philips, E. J. Rubin, and N. Perrimon (2005)
Science 309, 1251-1253
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