Automated genome-wide visual profiling of cellular proteins involved in HIV infection
| Autor: | Marc P. Windisch, Neil Emans, Nam Youl Kim, Ulf Nehrbass, Virginie Perrin, Seo Yeon Choi, Sungyong Jung, Auguste Genovesio, Fabrizio Mammano, Hi Chul Kim, Annette S Boese, Olivier Schwartz, Nicoletta Casartelli, Yong-Jun Kwon |
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| Přispěvatelé: | Institut Pasteur Korea - Institut Pasteur de Corée, Réseau International des Instituts Pasteur (RIIP), Yonsei University, Virus et Immunité, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), the work, conducted at institut pasteur Korea, was supported by Institut pasteur Korea and the Korean ministry of science andtechnology. the work conducted by olivier schwartz’s laboratory was supported by grants from agence nationale de recherche sur le sida (anrs), sidaction, cnrs, european community (fp7 contract 201412), and institut pasteur, there was no commercial support for this project., European Project: 201412,EC:FP7:HEALTH,FP7-HEALTH-2007-A,INEF(2008), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris] |
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Proteomics
[SDV]Life Sciences [q-bio] MESH: Antigens CD4 MESH: Automation Laboratory Virus Replication Biochemistry Jurkat cells Analytical Chemistry Small hairpin RNA Jurkat Cells 0302 clinical medicine RNA interference MESH: Jurkat Cells MESH: Microscopy Confocal MESH: Ku Autoantigen Protein Kinase C 0303 health sciences Microscopy Confocal MESH: HIV MESH: Proteomics Antigens Nuclear MESH: Gene Expression Regulation 3. Good health Housekeeping gene DNA-Binding Proteins High-content screening Gene Knockdown Techniques CD4 Antigens Molecular Medicine RNA Interference Biotechnology MESH: Cell Line Tumor Ribonuclease H MESH: RNA Interference Computational biology Biology 03 medical and health sciences Cell Line Tumor Gene silencing Humans Gene Ku Autoantigen MESH: Antigens Nuclear 030304 developmental biology Automation Laboratory MESH: Ribonuclease H MESH: Humans MESH: Virus Replication HIV Microarray Analysis Virology MESH: Protein Kinase C MESH: Gene Knockdown Techniques MESH: Microarray Analysis Viral replication Gene Expression Regulation MESH: HeLa Cells 030217 neurology & neurosurgery MESH: DNA-Binding Proteins HeLa Cells |
| Zdroj: | Journal of Biomolecular Screening Journal of Biomolecular Screening, 2011, 16 (9), pp.945-58. ⟨10.1177/1087057111415521⟩ Journal of Biomolecular Screening; Vol 16 Journal of Biomolecular Screening, SAGE Publications, 2011, 16 (9), pp.945-58. ⟨10.1177/1087057111415521⟩ |
| ISSN: | 1087-0571 |
| DOI: | 10.1177/1087057111415521⟩ |
| Popis: | International audience; Recent genome-wide RNAi screens have identified >842 human genes that affect the human immunodeficiency virus (HIV) cycle. The list of genes implicated in infection differs between screens, and there is minimal overlap. A reason for this variance is the interdependence of HIV infection and host cell function, producing a multitude of indirect or pleiotropic cellular effects affecting the viral infection during RNAi screening. To overcome this, the authors devised a 15-dimensional phenotypic profile to define the viral infection block induced by CD4 silencing in HeLa cells. They demonstrate that this phenotypic profile excludes nonspecific, RNAi-based side effects and viral replication defects mediated by silencing of housekeeping genes. To achieve statistical robustness, the authors used automatically annotated RNAi arrays for seven independent genome-wide RNAi screens. This identified 56 host genes, which reliably reproduced CD4-like phenotypes upon HIV infection. The factors include 11 known HIV interactors and 45 factors previously not associated with HIV infection. As proof of concept, the authors confirmed that silencing of PAK1, Ku70, and RNAseH2A impaired HIV replication in Jurkat cells. In summary, multidimensional, visual profiling can identify genes required for HIV infection. |
| Databáze: | OpenAIRE |
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