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Article: A genome-wide short hairpin RNA screening of Jurkat T-cells for human proteins contributing to productive HIV-1 replication

TitleA genome-wide short hairpin RNA screening of Jurkat T-cells for human proteins contributing to productive HIV-1 replication
Authors
Issue Date2009
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
Citation
Journal Of Biological Chemistry, 2009, v. 284 n. 29, p. 19463-19473 How to Cite?
AbstractShort interfering RNAs (siRNAs) have been used to inhibit HIV-1 replication. The durable inhibition of HIV-1 replication by RNA interference has been impeded, however by a high mutation rate when viral sequences art targeted and by cytotoxicity when cellular genes are knocked down. To identify cellular proteins that contribute to HIV-1 replication that coin be chronically silenced without significant cytotoxicity, we employed a shRNA library that targets 54,509 human transcripts. We used this library to select a comprehensive population of Jurkat T-cell clones, each expressing a single discrete shRNA. The Jurkat clones were then infected with HIV-1. Clones that survived viral infection represent moieties silenced for a human mRNA needed for virus replication, but whose chronic knockdown did not cause cytotoxicity. Overall, 252 individual jurkat mRNAs were identified. Twenty-two of these mRNAs were secondarily verified for their contributions to HIV-1 replication. Five mRNAs, NRF1, STXBP2, NCOA3, PRDM2, and EXOSCS, were studied for their effect on steps of the HIV-1 life cycle. We discuss the similarities and differences between our shRNA findings for HIV-1 using a spreading infection assay in human Jurkat T-cells and results from other investigators who used siRNA-based screenings in HeLa or 293T cells.
Persistent Identifierhttp://hdl.handle.net/10722/157553
ISSN
2015 Impact Factor: 4.258
2015 SCImago Journal Rankings: 3.151
ISI Accession Number ID
Funding AgencyGrant Number
NIAID
Office of the Director, NIH
Funding Information:

This work was supported, in whole or in part, by National Institutes of Health intramural research funds from NIAID. This work was also supported by the IATAP program from the Office of the Director, NIH.

References

 

DC FieldValueLanguage
dc.contributor.authorYeung, MLen_US
dc.contributor.authorHouzet, Len_US
dc.contributor.authorYedavalli, VSRKen_US
dc.contributor.authorJeang, KTen_US
dc.date.accessioned2012-08-08T08:51:12Z-
dc.date.available2012-08-08T08:51:12Z-
dc.date.issued2009en_US
dc.identifier.citationJournal Of Biological Chemistry, 2009, v. 284 n. 29, p. 19463-19473en_US
dc.identifier.issn0021-9258en_US
dc.identifier.urihttp://hdl.handle.net/10722/157553-
dc.description.abstractShort interfering RNAs (siRNAs) have been used to inhibit HIV-1 replication. The durable inhibition of HIV-1 replication by RNA interference has been impeded, however by a high mutation rate when viral sequences art targeted and by cytotoxicity when cellular genes are knocked down. To identify cellular proteins that contribute to HIV-1 replication that coin be chronically silenced without significant cytotoxicity, we employed a shRNA library that targets 54,509 human transcripts. We used this library to select a comprehensive population of Jurkat T-cell clones, each expressing a single discrete shRNA. The Jurkat clones were then infected with HIV-1. Clones that survived viral infection represent moieties silenced for a human mRNA needed for virus replication, but whose chronic knockdown did not cause cytotoxicity. Overall, 252 individual jurkat mRNAs were identified. Twenty-two of these mRNAs were secondarily verified for their contributions to HIV-1 replication. Five mRNAs, NRF1, STXBP2, NCOA3, PRDM2, and EXOSCS, were studied for their effect on steps of the HIV-1 life cycle. We discuss the similarities and differences between our shRNA findings for HIV-1 using a spreading infection assay in human Jurkat T-cells and results from other investigators who used siRNA-based screenings in HeLa or 293T cells.en_US
dc.languageengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/en_US
dc.relation.ispartofJournal of Biological Chemistryen_US
dc.subject.meshAntigens, Neoplasm - Geneticsen_US
dc.subject.meshAntigens, Surface - Geneticsen_US
dc.subject.meshCell Lineen_US
dc.subject.meshCell Proliferationen_US
dc.subject.meshDna-Binding Proteins - Geneticsen_US
dc.subject.meshExoribonucleases - Geneticsen_US
dc.subject.meshFlow Cytometryen_US
dc.subject.meshGene Libraryen_US
dc.subject.meshGenome, Humanen_US
dc.subject.meshHiv Core Protein P24 - Genetics - Metabolismen_US
dc.subject.meshHiv-1 - Genetics - Growth & Development - Metabolismen_US
dc.subject.meshHela Cellsen_US
dc.subject.meshHistone Acetyltransferases - Geneticsen_US
dc.subject.meshHistone-Lysine N-Methyltransferaseen_US
dc.subject.meshHumansen_US
dc.subject.meshJurkat Cellsen_US
dc.subject.meshMunc18 Proteins - Geneticsen_US
dc.subject.meshNuclear Proteins - Geneticsen_US
dc.subject.meshNuclear Receptor Coactivator 3en_US
dc.subject.meshNuclear Respiratory Factor 1 - Geneticsen_US
dc.subject.meshOligonucleotide Array Sequence Analysis - Methodsen_US
dc.subject.meshProteins - Geneticsen_US
dc.subject.meshRna, Messenger - Geneticsen_US
dc.subject.meshRna, Small Interfering - Geneticsen_US
dc.subject.meshTrans-Activators - Geneticsen_US
dc.subject.meshTranscription Factors - Geneticsen_US
dc.subject.meshVirus Replication - Geneticsen_US
dc.titleA genome-wide short hairpin RNA screening of Jurkat T-cells for human proteins contributing to productive HIV-1 replicationen_US
dc.typeArticleen_US
dc.identifier.emailYeung, ML:pmlyeung@hku.hken_US
dc.identifier.authorityYeung, ML=rp01402en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1074/jbc.M109.010033en_US
dc.identifier.pmid19460752-
dc.identifier.scopuseid_2-s2.0-67749148923en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-67749148923&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume284en_US
dc.identifier.issue29en_US
dc.identifier.spage19463en_US
dc.identifier.epage19473en_US
dc.identifier.isiWOS:000267908300038-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridYeung, ML=8350940900en_US
dc.identifier.scopusauthoridHouzet, L=7801516884en_US
dc.identifier.scopusauthoridYedavalli, VSRK=6603053420en_US
dc.identifier.scopusauthoridJeang, KT=7004824803en_US

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