Derivation of a novel SARS–coronavirus replicon cell line and its application for anti-SARS drug screening
Identifieur interne : 001554 ( Pmc/Curation ); précédent : 001553; suivant : 001555Derivation of a novel SARS–coronavirus replicon cell line and its application for anti-SARS drug screening
Auteurs : Feng Ge [Singapour] ; Yonghu Luo [République populaire de Chine] ; Pei Xiong Liew ; Eugene Hung [Singapour]Source :
- Virology [ 0042-6822 ] ; 2006.
Abstract
The severe acute respiratory syndrome (SARS) outbreak in 2002, which had a high morbidity rate and caused worldwide alarm, remains untreated today even though SARS was eventually isolated and controlled. Development and high-throughput screening of efficacious drugs is therefore critical. However, currently there remains a lack of such a safe system. Here, the generation and characterization of the first selectable, SARS–coronavirus (SARS–CoV)-based replicon cell line which can be used for screening is described. Partial SARS–CoV cDNAs and antibiotic resistance/reporter gene DNA were generated and assembled in vitro to produce the replicon transcription template, which was then transcribed in vitro to generate the replicon RNA. The latter was introduced into a mammalian cell line and the transfected cells were selected for by antibiotic application. For the antibiotic-resistant cell lines thus generated, the expression of reporter gene was ensured by continued monitoring using fluorescent microscopy and flow cytometry. The suitability of this replicon cell line in drug screening was demonstrated by testing the inhibitory effect of several existing drugs and the results demonstrate that the SARS–CoV replicon cell lines provide a safe tool for the identification of SARS–CoV replicase inhibitors. The replicon cell lines thus developed can be applied to high-throughput screening for anti-SARS drugs without the need to grow infectious SARS–CoV.
Url:
DOI: 10.1016/j.virol.2006.10.016
PubMed: 17098272
PubMed Central: 7103326
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Pei Xiong Liew<affiliation><nlm:aff id="aff3">Department of Haematology, Singapore General Hospital, Singapore Health Research Facilities, 7 Hospital Drive, Block A, #02-05, Singapore 169611, Republic of Singapore</nlm:aff><wicri:noCountry code="subfield">Republic of Singapore</wicri:noCountry></affiliation>Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Derivation of a novel SARS–coronavirus replicon cell line and its application for anti-SARS drug screening</title><author><name sortKey="Ge, Feng" sort="Ge, Feng" uniqKey="Ge F" first="Feng" last="Ge">Feng Ge</name><affiliation wicri:level="4"><nlm:aff id="aff1">Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Block MD4, 5 Science Drive 2 Singapore 117597, Republic of Singapore</nlm:aff><orgName type="university">Université nationale de Singapour</orgName><country>Singapour</country></affiliation></author><author><name sortKey="Luo, Yonghu" sort="Luo, Yonghu" uniqKey="Luo Y" first="Yonghu" last="Luo">Yonghu Luo</name><affiliation wicri:level="1"><nlm:aff id="aff2">Guangdong Institute of Microbiology, 100 Xianlie Central Road, Guangzhou, Guangdong, 510070, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Guangdong Institute of Microbiology, 100 Xianlie Central Road, Guangzhou, Guangdong, 510070</wicri:regionArea></affiliation></author><author><name sortKey="Liew, Pei Xiong" sort="Liew, Pei Xiong" uniqKey="Liew P" first="Pei Xiong" last="Liew">Pei Xiong Liew</name><affiliation><nlm:aff id="aff3">Department of Haematology, Singapore General Hospital, Singapore Health Research Facilities, 7 Hospital Drive, Block A, #02-05, Singapore 169611, Republic of Singapore</nlm:aff><wicri:noCountry code="subfield">Republic of Singapore</wicri:noCountry></affiliation></author><author><name sortKey="Hung, Eugene" sort="Hung, Eugene" uniqKey="Hung E" first="Eugene" last="Hung">Eugene Hung</name><affiliation wicri:level="4"><nlm:aff id="aff1">Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Block MD4, 5 Science Drive 2 Singapore 117597, Republic of Singapore</nlm:aff><orgName type="university">Université nationale de Singapour</orgName><country>Singapour</country></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">17098272</idno><idno type="pmc">7103326</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7103326</idno><idno type="RBID">PMC:7103326</idno><idno type="doi">10.1016/j.virol.2006.10.016</idno><date when="2006">2006</date><idno type="wicri:Area/Pmc/Corpus">001554</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001554</idno><idno type="wicri:Area/Pmc/Curation">001554</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">001554</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Derivation of a novel SARS–coronavirus replicon cell line and its application for anti-SARS drug screening</title><author><name sortKey="Ge, Feng" sort="Ge, Feng" uniqKey="Ge F" first="Feng" last="Ge">Feng Ge</name><affiliation wicri:level="4"><nlm:aff id="aff1">Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Block MD4, 5 Science Drive 2 Singapore 117597, Republic of Singapore</nlm:aff><orgName type="university">Université nationale de Singapour</orgName><country>Singapour</country></affiliation></author><author><name sortKey="Luo, Yonghu" sort="Luo, Yonghu" uniqKey="Luo Y" first="Yonghu" last="Luo">Yonghu Luo</name><affiliation wicri:level="1"><nlm:aff id="aff2">Guangdong Institute of Microbiology, 100 Xianlie Central Road, Guangzhou, Guangdong, 510070, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Guangdong Institute of Microbiology, 100 Xianlie Central Road, Guangzhou, Guangdong, 510070</wicri:regionArea></affiliation></author><author><name sortKey="Liew, Pei Xiong" sort="Liew, Pei Xiong" uniqKey="Liew P" first="Pei Xiong" last="Liew">Pei Xiong Liew</name><affiliation><nlm:aff id="aff3">Department of Haematology, Singapore General Hospital, Singapore Health Research Facilities, 7 Hospital Drive, Block A, #02-05, Singapore 169611, Republic of Singapore</nlm:aff><wicri:noCountry code="subfield">Republic of Singapore</wicri:noCountry></affiliation></author><author><name sortKey="Hung, Eugene" sort="Hung, Eugene" uniqKey="Hung E" first="Eugene" last="Hung">Eugene Hung</name><affiliation wicri:level="4"><nlm:aff id="aff1">Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Block MD4, 5 Science Drive 2 Singapore 117597, Republic of Singapore</nlm:aff><orgName type="university">Université nationale de Singapour</orgName><country>Singapour</country></affiliation></author></analytic><series><title level="j">Virology</title><idno type="ISSN">0042-6822</idno><idno type="eISSN">1096-0341</idno><imprint><date when="2006">2006</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The severe acute respiratory syndrome (SARS) outbreak in 2002, which had a high morbidity rate and caused worldwide alarm, remains untreated today even though SARS was eventually isolated and controlled. Development and high-throughput screening of efficacious drugs is therefore critical. However, currently there remains a lack of such a safe system. Here, the generation and characterization of the first selectable, SARS–coronavirus (SARS–CoV)-based replicon cell line which can be used for screening is described. Partial SARS–CoV cDNAs and antibiotic resistance/reporter gene DNA were generated and assembled in vitro to produce the replicon transcription template, which was then transcribed in vitro to generate the replicon RNA. The latter was introduced into a mammalian cell line and the transfected cells were selected for by antibiotic application. For the antibiotic-resistant cell lines thus generated, the expression of reporter gene was ensured by continued monitoring using fluorescent microscopy and flow cytometry. The suitability of this replicon cell line in drug screening was demonstrated by testing the inhibitory effect of several existing drugs and the results demonstrate that the SARS–CoV replicon cell lines provide a safe tool for the identification of SARS–CoV replicase inhibitors. The replicon cell lines thus developed can be applied to high-throughput screening for anti-SARS drugs without the need to grow infectious SARS–CoV.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Almazan, F" uniqKey="Almazan F">F. Almazan</name></author><author><name sortKey="Galan, C" uniqKey="Galan C">C. Galan</name></author><author><name sortKey="Enjuanes, L" uniqKey="Enjuanes L">L. Enjuanes</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Alonso, S" uniqKey="Alonso S">S. Alonso</name></author><author><name sortKey="Izeta, A" uniqKey="Izeta A">A. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Virology</journal-id><journal-id journal-id-type="iso-abbrev">Virology</journal-id><journal-title-group><journal-title>Virology</journal-title></journal-title-group><issn pub-type="ppub">0042-6822</issn><issn pub-type="epub">1096-0341</issn><publisher><publisher-name>Elsevier Inc.</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">17098272</article-id><article-id pub-id-type="pmc">7103326</article-id><article-id pub-id-type="publisher-id">S0042-6822(06)00741-0</article-id><article-id pub-id-type="doi">10.1016/j.virol.2006.10.016</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Derivation of a novel SARS–coronavirus replicon cell line and its application for anti-SARS drug screening</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Ge</surname><given-names>Feng</given-names></name><email>ge.feng@singhealth.com.sg</email><xref rid="aff1" ref-type="aff">a</xref><xref rid="cor1" ref-type="corresp">⁎</xref></contrib><contrib contrib-type="author"><name><surname>Luo</surname><given-names>Yonghu</given-names></name><xref rid="aff2" ref-type="aff">b</xref></contrib><contrib contrib-type="author"><name><surname>Liew</surname><given-names>Pei Xiong</given-names></name><xref rid="aff3" ref-type="aff">c</xref></contrib><contrib contrib-type="author"><name><surname>Hung</surname><given-names>Eugene</given-names></name><xref rid="aff1" ref-type="aff">a</xref></contrib></contrib-group><aff id="aff1"><label>a</label>Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Block MD4, 5 Science Drive 2 Singapore 117597, Republic of Singapore</aff><aff id="aff2"><label>b</label>Guangdong Institute of Microbiology, 100 Xianlie Central Road, Guangzhou, Guangdong, 510070, PR China</aff><aff id="aff3"><label>c</label>Department of Haematology, Singapore General Hospital, Singapore Health Research Facilities, 7 Hospital Drive, Block A, #02-05, Singapore 169611, Republic of Singapore</aff><author-notes><corresp id="cor1"><label>⁎</label>Corresponding author. Present address: Department of Clinical Research, SingHealth Research Facilities, Singapore General Hospital, Singapore 169611, Republic of Singapore. Fax: +65 6321 3606. <email>ge.feng@singhealth.com.sg</email></corresp></author-notes><pub-date pub-type="pmc-release"><day>13</day><month>11</month><year>2006</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><day>30</day><month>3</month><year>2007</year></pub-date><pub-date pub-type="epub"><day>13</day><month>11</month><year>2006</year></pub-date><volume>360</volume><issue>1</issue><fpage>150</fpage><lpage>158</lpage><history><date date-type="received"><day>9</day><month>8</month><year>2006</year></date><date date-type="rev-recd"><day>30</day><month>8</month><year>2006</year></date><date date-type="accepted"><day>1</day><month>10</month><year>2006</year></date></history><permissions><copyright-statement>Copyright © 2006 Elsevier Inc. All rights reserved.</copyright-statement><copyright-year>2006</copyright-year><copyright-holder>Elsevier Inc.</copyright-holder><license><license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p></license></permissions><abstract><p>The severe acute respiratory syndrome (SARS) outbreak in 2002, which had a high morbidity rate and caused worldwide alarm, remains untreated today even though SARS was eventually isolated and controlled. Development and high-throughput screening of efficacious drugs is therefore critical. However, currently there remains a lack of such a safe system. Here, the generation and characterization of the first selectable, SARS–coronavirus (SARS–CoV)-based replicon cell line which can be used for screening is described. Partial SARS–CoV cDNAs and antibiotic resistance/reporter gene DNA were generated and assembled in vitro to produce the replicon transcription template, which was then transcribed in vitro to generate the replicon RNA. The latter was introduced into a mammalian cell line and the transfected cells were selected for by antibiotic application. For the antibiotic-resistant cell lines thus generated, the expression of reporter gene was ensured by continued monitoring using fluorescent microscopy and flow cytometry. The suitability of this replicon cell line in drug screening was demonstrated by testing the inhibitory effect of several existing drugs and the results demonstrate that the SARS–CoV replicon cell lines provide a safe tool for the identification of SARS–CoV replicase inhibitors. The replicon cell lines thus developed can be applied to high-throughput screening for anti-SARS drugs without the need to grow infectious SARS–CoV.</p></abstract><kwd-group><title>Keywords</title><kwd>SARS</kwd><kwd>Coronavirus</kwd><kwd>SARS–CoV</kwd><kwd>Reverse genetics</kwd><kwd>Replicon</kwd><kwd>Drug screening</kwd></kwd-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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