Inhibition of replication and infection of severe acute respiratory syndrome-associated coronavirus with plasmid-mediated interference RNA.
Identifieur interne : 004782 ( Main/Exploration ); précédent : 004781; suivant : 004783Inhibition of replication and infection of severe acute respiratory syndrome-associated coronavirus with plasmid-mediated interference RNA.
Auteurs : Bing Ni [République populaire de Chine] ; Xinfu Shi ; Yang Li ; Wenda Gao ; Xiliang Wang ; Yuzhang WuSource :
- Antiviral therapy [ 1359-6535 ] ; 2005.
Descripteurs français
- KwdFr :
- Animaux, Cellules Vero, Effet cytopathogène viral (génétique), Interférence par ARN, Petit ARN interférent (pharmacologie), Plasmides (génétique), Protéines virales non structurales (génétique), Régulation de l'expression des gènes viraux (physiologie), Réplication virale (génétique), Virus du SRAS (génétique), Virus du SRAS (physiologie).
- MESH :
- génétique : Effet cytopathogène viral, Plasmides, Protéines virales non structurales, Réplication virale, Virus du SRAS.
- pharmacologie : Petit ARN interférent.
- physiologie : Régulation de l'expression des gènes viraux, Virus du SRAS.
- Animaux, Cellules Vero, Interférence par ARN.
English descriptors
- KwdEn :
- Animals, Chlorocebus aethiops, Cytopathogenic Effect, Viral (genetics), Gene Expression Regulation, Viral (physiology), Plasmids (genetics), RNA Interference, RNA, Small Interfering (pharmacology), SARS Virus (genetics), SARS Virus (physiology), Vero Cells, Viral Nonstructural Proteins (genetics), Virus Replication (genetics).
- MESH :
- chemical , genetics : Viral Nonstructural Proteins.
- chemical , pharmacology : RNA, Small Interfering.
- genetics : Cytopathogenic Effect, Viral, Plasmids, SARS Virus, Virus Replication.
- physiology : Gene Expression Regulation, Viral, SARS Virus.
- Animals, Chlorocebus aethiops, RNA Interference, Vero Cells.
Abstract
Severe acute respiratory syndrome (SARS) is a newly emerged infectious disease caused by a novel coronavirus (SARS-CoV), which spread to over 30 countries in early 2003. Until recently, no specific vaccines and effective drugs have been available to protect patients from infection by this virus. To exploit a new strategy to fight this disease, we investigated the effect of interference RNA (RNAi) on the virus infection and replication with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT), plaque-forming, Western-blot and real-time PCR assays. Results showed that the plasmid-derived siRNAs targeting the non-structural protein 1 (NSP1) sequence of the SARS-CoV genome could specifically inhibit the expression of the NSP1 sequence and effectively suppress the replication and propagation of SARS-CoV in cultured Vero E6 cell lines. The expression of the Spike and Nucleoprotein genes of SARS-CoV at mRNA and protein levels in small interfering (si)RNA-expressing cells was significantly less than that in controls when analysed with PCR and Western-blot assays, 3 days post SARS-CoV infection. Our study provides strong evidence that the NSP1 sequence in the SARS-CoV genome is a valid target for RNAi and the effect of the siRNAs probably mainly resulted from effects on global reduction of subgenome synthesis and subsequent protein expression of SARS-CoV.
PubMed: 16038478
Affiliations:
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uniqKey="Gao W" first="Wenda" last="Gao">Wenda Gao</name></author><author><name sortKey="Wang, Xiliang" sort="Wang, Xiliang" uniqKey="Wang X" first="Xiliang" last="Wang">Xiliang Wang</name></author><author><name sortKey="Wu, Yuzhang" sort="Wu, Yuzhang" uniqKey="Wu Y" first="Yuzhang" last="Wu">Yuzhang Wu</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:16038478</idno><idno type="pmid">16038478</idno><idno type="wicri:Area/PubMed/Corpus">002627</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002627</idno><idno type="wicri:Area/PubMed/Curation">002627</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002627</idno><idno type="wicri:Area/PubMed/Checkpoint">002650</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002650</idno><idno type="wicri:Area/Ncbi/Merge">001074</idno><idno 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Chongqing</wicri:regionArea><wicri:noRegion>Chongqing</wicri:noRegion></affiliation></author><author><name sortKey="Shi, Xinfu" sort="Shi, Xinfu" uniqKey="Shi X" first="Xinfu" last="Shi">Xinfu Shi</name></author><author><name sortKey="Li, Yang" sort="Li, Yang" uniqKey="Li Y" first="Yang" last="Li">Yang Li</name></author><author><name sortKey="Gao, Wenda" sort="Gao, Wenda" uniqKey="Gao W" first="Wenda" last="Gao">Wenda Gao</name></author><author><name sortKey="Wang, Xiliang" sort="Wang, Xiliang" uniqKey="Wang X" first="Xiliang" last="Wang">Xiliang Wang</name></author><author><name sortKey="Wu, Yuzhang" sort="Wu, Yuzhang" uniqKey="Wu Y" first="Yuzhang" last="Wu">Yuzhang Wu</name></author></analytic><series><title level="j">Antiviral therapy</title><idno type="ISSN">1359-6535</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Cytopathogenic Effect, Viral (genetics)</term><term>Gene Expression Regulation, Viral (physiology)</term><term>Plasmids (genetics)</term><term>RNA Interference</term><term>RNA, Small Interfering (pharmacology)</term><term>SARS Virus (genetics)</term><term>SARS Virus (physiology)</term><term>Vero Cells</term><term>Viral Nonstructural Proteins (genetics)</term><term>Virus Replication (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>Effet cytopathogène viral (génétique)</term><term>Interférence par ARN</term><term>Petit ARN interférent (pharmacologie)</term><term>Plasmides (génétique)</term><term>Protéines virales non structurales (génétique)</term><term>Régulation de l'expression des gènes viraux (physiologie)</term><term>Réplication virale (génétique)</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>RNA, Small Interfering</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Cytopathogenic Effect, Viral</term><term>Plasmids</term><term>SARS Virus</term><term>Virus Replication</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Effet cytopathogène viral</term><term>Plasmides</term><term>Protéines virales non structurales</term><term>Réplication virale</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Petit ARN interférent</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Régulation de l'expression des gènes viraux</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Gene Expression Regulation, Viral</term><term>SARS Virus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>RNA Interference</term><term>Vero Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>Interférence par ARN</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) is a newly emerged infectious disease caused by a novel coronavirus (SARS-CoV), which spread to over 30 countries in early 2003. Until recently, no specific vaccines and effective drugs have been available to protect patients from infection by this virus. To exploit a new strategy to fight this disease, we investigated the effect of interference RNA (RNAi) on the virus infection and replication with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT), plaque-forming, Western-blot and real-time PCR assays. Results showed that the plasmid-derived siRNAs targeting the non-structural protein 1 (NSP1) sequence of the SARS-CoV genome could specifically inhibit the expression of the NSP1 sequence and effectively suppress the replication and propagation of SARS-CoV in cultured Vero E6 cell lines. The expression of the Spike and Nucleoprotein genes of SARS-CoV at mRNA and protein levels in small interfering (si)RNA-expressing cells was significantly less than that in controls when analysed with PCR and Western-blot assays, 3 days post SARS-CoV infection. Our study provides strong evidence that the NSP1 sequence in the SARS-CoV genome is a valid target for RNAi and the effect of the siRNAs probably mainly resulted from effects on global reduction of subgenome synthesis and subsequent protein expression of SARS-CoV.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><noCountry><name sortKey="Gao, Wenda" sort="Gao, Wenda" uniqKey="Gao W" first="Wenda" last="Gao">Wenda Gao</name><name sortKey="Li, Yang" sort="Li, Yang" uniqKey="Li Y" first="Yang" last="Li">Yang Li</name><name sortKey="Shi, Xinfu" sort="Shi, Xinfu" uniqKey="Shi X" first="Xinfu" last="Shi">Xinfu Shi</name><name sortKey="Wang, Xiliang" sort="Wang, Xiliang" uniqKey="Wang X" first="Xiliang" last="Wang">Xiliang Wang</name><name sortKey="Wu, Yuzhang" sort="Wu, Yuzhang" uniqKey="Wu Y" first="Yuzhang" last="Wu">Yuzhang Wu</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Ni, Bing" sort="Ni, Bing" uniqKey="Ni B" first="Bing" last="Ni">Bing Ni</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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