title: Probing the virus host interaction in high containment: an approach using pooled short hairpin RNA.
abstract: The study of viruses in high containment offers unique challenges for technology-intense approaches. These approaches include high-throughput screening for small-molecule antivirals and genetic perturbation-based screens for host factors required for viral replication. Here, we describe the use of whole-genome scale pooled short hairpin RNA (shRNA) libraries to screen for host factors necessary for viral infection at BSL2, and the transition of this technique into the BSL4 environment. Pooled screening provides a unique way to circumvent many of the technological challenges associated with other high-throughput screening approaches in high containment. Our pooled screening approach identified host factors involved in the replication of orthopoxviruses (Vaccinia and Monkeypox) and filoviruses (Ebola and Marburg) under conditions that enable straightforward screen-to-follow-up approaches.
authors: 1 National Emerging Infectious Diseases Laboratory, Boston University School of Medicine , Boston, Massachusetts.
authors: Claire Marie
authors: CM
authors: Filone
authors: Ken
authors: K
authors: Dower
authors: Glenn S
authors: GS
authors: Cowley
authors: Lisa E
authors: LE
authors: Hensley
authors: John H
authors: JH
authors: Connor
mesh_terms: D001681:Biological Assay / Q000379:methods; D003264:Containment of Biohazards; D015698:Genomic Library; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D054884:Host-Pathogen Interactions / Q000235:genetics*; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics*; D014615:Vaccinia / Q000821:virology*; D014616:Vaccinia virus / Q000502:physiology*; D016662:Virus Integration / Q000502:physiology
chemical_list: D034741:RNA, Small Interfering
references: Nature. 2010 Feb 11;463(7282):818-22
references: 20081832
references: Cell. 2010 Apr 30;141(3):497-508
references: 20434987
references: Proc Natl Acad Sci U S A. 2010 Jun 22;107(25):11513-8
references: 20534518
references: Mol Biol Rep. 2010 Jul;37(6):2863-70
references: 19798586
references: J Biomol Screen. 2010 Aug;15(7):755-65
references: 20639507
references: Lancet. 2011 Mar 5;377(9768):849-62
references: 21084112
references: Antiviral Res. 2011 Jul;91(1):72-80
references: 21569797
references: Proc Natl Acad Sci U S A. 2011 Jul 26;108(30):12372-7
references: 21746896
references: J Virol. 2011 Oct;85(19):9899-908
references: 21795349
references: Nature. 2011 Sep 15;477(7364):344-8
references: 21866101
references: Nature. 2011 Sep 15;477(7364):340-3
references: 21866103
references: Trends Microbiol. 2011 Oct;19(10):501-8
references: 21855347
references: J Virol. 2012 Mar;86(5):2632-40
references: 22205744
references: J Virol. 2012 Mar;86(6):3284-92
references: 22238307
references: Virol J. 2012;9:5
references: 22225589
references: Mol Syst Biol. 2012;8:579
references: 22531119
references: Curr Opin Virol. 2012 Dec;2(6):784-92
references: 23122855
references: Antiviral Res. 2013 Sep;99(3):207-13
references: 23751367
references: Science. 2014 Jan 3;343(6166):80-4
references: 24336569
references: PLoS Pathog. 2014 Feb;10(2):e1003904
references: 24516381
references: Antiviral Res. 2014 Jun;106:86-94
references: 24713118
references: Lancet Infect Dis. 2014 Nov;14(11):1034-5
references: 25282665
references: Cytogenet Genome Res. 2002;97(1-2):72-5
references: 12438742
references: Biochem Biophys Res Commun. 2003 Nov 14;311(2):518-24
references: 14592447
references: Nature. 2004 Mar 25;428(6981):431-7
references: 15042092
references: J Virol. 1996 Oct;70(10):6909-21
references: 8794334
references: Virology. 2005 Feb 5;332(1):20-7
references: 15661137
references: Science. 2005 Jun 10;308(5728):1643-5
references: 15831716
references: Cell. 2005 Sep 9;122(5):735-49
references: 16143105
references: Nat Genet. 2005 Nov;37(11):1281-8
references: 16200065
references: Cell. 2006 Mar 24;124(6):1283-98
references: 16564017
references: Nat Rev Genet. 2006 May;7(5):373-84
references: 16607398
references: Nat Chem Biol. 2007 Aug;3(8):466-79
references: 17637779
references: J Biol Chem. 2007 Aug 17;282(33):23878-91
references: 17556371
references: RNA. 2007 Oct;13(10):1765-74
references: 17698642
references: J Infect Dis. 2007 Nov 15;196 Suppl 2:S313-22
references: 17940966
references: Nature. 2008 Mar 20;452(7185):323-8
references: 18272964
references: Proc Natl Acad Sci U S A. 2008 Dec 23;105(51):20380-5
references: 19091943
references: Curr Opin Cell Biol. 2009 Aug;21(4):543-51
references: 19577915