SrasV1, PascalFrancis, Corpus, bibRecord, 000244

High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery

Identifieur interne : 000244 ( PascalFrancis/Corpus ); précédent : 000243; suivant : 000245

High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery

Auteurs : FENG GE ; SHENG XIONG ; Fu-Sen Lin ; Zhi-Ping Zhang ; Xian-En Zhang

Source :

Antiviral research [ 0166-3542 ] ; 2008.

RBID : Pascal:08-0536390

Descripteurs français

Pascal (Inist)
- Criblage haut débit, Virus syndrome respiratoire aigu sévère, Recherche et développement, Médicament, Protéine fluorescente verte, Réplicon, Expression génique, Antiviral, Analyse à haut débit.

English descriptors

KwdEn :
- Antiviral, Drug, Gene expression, Green fluorescent protein, High throughput screening, Replicon, Research and development, Severe acute respiratory syndrome virus.

Abstract

The causative agent of severe acute respiratory syndrome (SARS) has been identified as a novel coronavirus, SARS-CoV. The development of rapid screening assays is essential for antiviral drug discovery. By using a cell line expressing a SARS-CoV subgenomic replicon, we developed a high-throughput assay and used it to screen small molecule compounds for inhibitors of SARS-CoV replication in the absence of live virus. The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses. Based on this assay system, we screened 7035 small molecule compounds from which we identified 7 compounds with anti-SARS-CoV activity. We demonstrate that the compounds inhibited SARS-CoV replication-dependent GFP expression in the replicon cells and reduced SARS-CoV viral protein accumulation and viral RNA copy number in the replicon cells. In a SARS-CoV plaque reduction assay, these compounds were confirmed to have antiviral activity. The target of one of the hit compounds, C12344, was validated by the generation of resistant replicon cells and the identification of the mutations conferring the resistant phenotype. These compounds should be valuable for developing anti-SARS therapeutic drugs as well as research tools to study the mechanism of SARS-CoV replication.

Notice en format standard (ISO 2709)

Pour connaître la documentation sur le format Inist Standard.

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A11	`03`	`1`		`@1 LIN (Fu-Sen)`
A11	`04`	`1`		`@1 ZHANG (Zhi-Ping)`
A11	`05`	`1`		`@1 ZHANG (Xian-En)`
A14	`01`			`@1 Institute of Life and Health Engineering,Jinan University @2 Guangzhou, Guangdong 510632 @3 CHN @Z 1 aut. @Z 2 aut.`
A14	`02`			`@1 Division of Research, Singapore General Hospital, Singapore Health Research Facilities, 7 Hospital Drive, Block A, #02-05 @2 Singapore 169611 @3 SGP @Z 3 aut.`
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C01	`01`		`ENG`	@0 The causative agent of severe acute respiratory syndrome (SARS) has been identified as a novel coronavirus, SARS-CoV. The development of rapid screening assays is essential for antiviral drug discovery. By using a cell line expressing a SARS-CoV subgenomic replicon, we developed a high-throughput assay and used it to screen small molecule compounds for inhibitors of SARS-CoV replication in the absence of live virus. The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses. Based on this assay system, we screened 7035 small molecule compounds from which we identified 7 compounds with anti-SARS-CoV activity. We demonstrate that the compounds inhibited SARS-CoV replication-dependent GFP expression in the replicon cells and reduced SARS-CoV viral protein accumulation and viral RNA copy number in the replicon cells. In a SARS-CoV plaque reduction assay, these compounds were confirmed to have antiviral activity. The target of one of the hit compounds, C12344, was validated by the generation of resistant replicon cells and the identification of the mutations conferring the resistant phenotype. These compounds should be valuable for developing anti-SARS therapeutic drugs as well as research tools to study the mechanism of SARS-CoV replication.
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C03	`06`	`X`	`SPA`	`@0 Replicón @5 06`
C03	`07`	`X`	`FRE`	`@0 Expression génique @5 07`
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C03	`07`	`X`	`SPA`	`@0 Expresión genética @5 07`
C03	`08`	`X`	`FRE`	`@0 Antiviral @5 09`
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Format Inist (serveur)

NO :	PASCAL 08-0536390 INIST
ET :	High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery
AU :	FENG GE; SHENG XIONG; LIN (Fu-Sen); ZHANG (Zhi-Ping); ZHANG (Xian-En)
AF :	Institute of Life and Health Engineering,Jinan University/Guangzhou, Guangdong 510632/Chine (1 aut., 2 aut.); Division of Research, Singapore General Hospital, Singapore Health Research Facilities, 7 Hospital Drive, Block A, #02-05/Singapore 169611/Singapour (3 aut.); State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences/Wuhan 430071/Chine (4 aut., 5 aut.)
DT :	Publication en série; Niveau analytique
SO :	Antiviral research; ISSN 0166-3542; Coden ARSRDR; Pays-Bas; Da. 2008; Vol. 80; No. 2; Pp. 107-113; Bibl. 3/4 p.
LA :	Anglais
EA :	The causative agent of severe acute respiratory syndrome (SARS) has been identified as a novel coronavirus, SARS-CoV. The development of rapid screening assays is essential for antiviral drug discovery. By using a cell line expressing a SARS-CoV subgenomic replicon, we developed a high-throughput assay and used it to screen small molecule compounds for inhibitors of SARS-CoV replication in the absence of live virus. The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses. Based on this assay system, we screened 7035 small molecule compounds from which we identified 7 compounds with anti-SARS-CoV activity. We demonstrate that the compounds inhibited SARS-CoV replication-dependent GFP expression in the replicon cells and reduced SARS-CoV viral protein accumulation and viral RNA copy number in the replicon cells. In a SARS-CoV plaque reduction assay, these compounds were confirmed to have antiviral activity. The target of one of the hit compounds, C12344, was validated by the generation of resistant replicon cells and the identification of the mutations conferring the resistant phenotype. These compounds should be valuable for developing anti-SARS therapeutic drugs as well as research tools to study the mechanism of SARS-CoV replication.
CC :	002B02S05
FD :	Criblage haut débit; Virus syndrome respiratoire aigu sévère; Recherche et développement; Médicament; Protéine fluorescente verte; Réplicon; Expression génique; Antiviral; Analyse à haut débit
FG :	Coronavirus; Coronaviridae; Nidovirales; Virus
ED :	High throughput screening; Severe acute respiratory syndrome virus; Research and development; Drug; Green fluorescent protein; Replicon; Gene expression; Antiviral
EG :	Coronavirus; Coronaviridae; Nidovirales; Virus
SD :	Cribado alta productividad; Severe acute respiratory syndrome virus; Investigación desarrollo; Medicamento; Proteína fluorescente verde; Replicón; Expresión genética; Antiviral
LO :	INIST-18839.354000184282020040
ID :	08-0536390

Links to Exploration step

Pascal:08-0536390

Le document en format XML

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<server><NO>PASCAL 08-0536390 INIST</NO>
<ET>High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery</ET>
<AU>FENG GE; SHENG XIONG; LIN (Fu-Sen); ZHANG (Zhi-Ping); ZHANG (Xian-En)</AU>
<AF>Institute of Life and Health Engineering,Jinan University/Guangzhou, Guangdong 510632/Chine (1 aut., 2 aut.); Division of Research, Singapore General Hospital, Singapore Health Research Facilities, 7 Hospital Drive, Block A, #02-05/Singapore 169611/Singapour (3 aut.); State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences/Wuhan 430071/Chine (4 aut., 5 aut.)</AF>
<DT>Publication en série; Niveau analytique</DT>
<SO>Antiviral research; ISSN 0166-3542; Coden ARSRDR; Pays-Bas; Da. 2008; Vol. 80; No. 2; Pp. 107-113; Bibl. 3/4 p.</SO>
<LA>Anglais</LA>
<EA>The causative agent of severe acute respiratory syndrome (SARS) has been identified as a novel coronavirus, SARS-CoV. The development of rapid screening assays is essential for antiviral drug discovery. By using a cell line expressing a SARS-CoV subgenomic replicon, we developed a high-throughput assay and used it to screen small molecule compounds for inhibitors of SARS-CoV replication in the absence of live virus. The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses. Based on this assay system, we screened 7035 small molecule compounds from which we identified 7 compounds with anti-SARS-CoV activity. We demonstrate that the compounds inhibited SARS-CoV replication-dependent GFP expression in the replicon cells and reduced SARS-CoV viral protein accumulation and viral RNA copy number in the replicon cells. In a SARS-CoV plaque reduction assay, these compounds were confirmed to have antiviral activity. The target of one of the hit compounds, C12344, was validated by the generation of resistant replicon cells and the identification of the mutations conferring the resistant phenotype. These compounds should be valuable for developing anti-SARS therapeutic drugs as well as research tools to study the mechanism of SARS-CoV replication.</EA>
<CC>002B02S05</CC>
<FD>Criblage haut débit; Virus syndrome respiratoire aigu sévère; Recherche et développement; Médicament; Protéine fluorescente verte; Réplicon; Expression génique; Antiviral; Analyse à haut débit</FD>
<FG>Coronavirus; Coronaviridae; Nidovirales; Virus</FG>
<ED>High throughput screening; Severe acute respiratory syndrome virus; Research and development; Drug; Green fluorescent protein; Replicon; Gene expression; Antiviral</ED>
<EG>Coronavirus; Coronaviridae; Nidovirales; Virus</EG>
<SD>Cribado alta productividad; Severe acute respiratory syndrome virus; Investigación desarrollo; Medicamento; Proteína fluorescente verde; Replicón; Expresión genética; Antiviral</SD>
<LO>INIST-18839.354000184282020040</LO>
<ID>08-0536390</ID>
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High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery

High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery

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