Reverse genetics of SARS-related coronavirus using vaccinia virus-based recombination.
Identifieur interne : 001C70 ( Main/Exploration ); précédent : 001C69; suivant : 001C71Reverse genetics of SARS-related coronavirus using vaccinia virus-based recombination.
Auteurs : Sjoerd H E. Van Den Worm [Pays-Bas] ; Klara Kristin Eriksson ; Jessika C. Zevenhoven ; Friedemann Weber ; Roland Züst ; Thomas Kuri ; Ronald Dijkman ; Guohui Chang ; Stuart G. Siddell ; Eric J. Snijder ; Volker Thiel ; Andrew D. DavidsonSource :
- PloS one [ 1932-6203 ] ; 2012.
Descripteurs français
- KwdFr :
- ADN complémentaire, Analyse de séquence d'ADN, Animaux, Cellules dendritiques (virologie), Clonage moléculaire, Données de séquences moléculaires, Génome viral, Humains, Lignée cellulaire, Méthode des plages virales, Ordre des gènes, Recombinaison génétique, Régulation de l'expression des gènes viraux, Réplication virale, Virus de la vaccine (génétique), Virus du SRAS (croissance et développement), Virus du SRAS (génétique), Virus recombinants (génétique).
- MESH :
- croissance et développement : Virus du SRAS.
- génétique : Virus de la vaccine, Virus du SRAS, Virus recombinants.
- virologie : Cellules dendritiques.
- ADN complémentaire, Analyse de séquence d'ADN, Animaux, Clonage moléculaire, Données de séquences moléculaires, Génome viral, Humains, Lignée cellulaire, Méthode des plages virales, Ordre des gènes, Recombinaison génétique, Régulation de l'expression des gènes viraux, Réplication virale.
English descriptors
- KwdEn :
- Animals, Cell Line, Chlorocebus aethiops, Cloning, Molecular, DNA, Complementary, Dendritic Cells (virology), Gene Expression Regulation, Viral, Gene Order, Genome, Viral, Humans, Molecular Sequence Data, Reassortant Viruses (genetics), Recombination, Genetic, SARS Virus (genetics), SARS Virus (growth & development), Sequence Analysis, DNA, Vaccinia virus (genetics), Viral Plaque Assay, Virus Replication.
- MESH :
- chemical : DNA, Complementary.
- genetics : Reassortant Viruses, SARS Virus, Vaccinia virus.
- growth & development : SARS Virus.
- virology : Dendritic Cells.
- Animals, Cell Line, Chlorocebus aethiops, Cloning, Molecular, Gene Expression Regulation, Viral, Gene Order, Genome, Viral, Humans, Molecular Sequence Data, Recombination, Genetic, Sequence Analysis, DNA, Viral Plaque Assay, Virus Replication.
Abstract
Severe acute respiratory syndrome (SARS) is a zoonotic disease caused by SARS-related coronavirus (SARS-CoV) that emerged in 2002 to become a global health concern. Although the original outbreak was controlled by classical public health measures, there is a real risk that another SARS-CoV could re-emerge from its natural reservoir, either in its original form or as a more virulent or pathogenic strain; in which case, the virus would be difficult to control in the absence of any effective antiviral drugs or vaccines. Using the well-studied SARS-CoV isolate HKU-39849, we developed a vaccinia virus-based SARS-CoV reverse genetic system that is both robust and biosafe. The SARS-CoV genome was cloned in separate vaccinia virus vectors, (vSARS-CoV-5prime and vSARS-CoV-3prime) as two cDNAs that were subsequently ligated to create a genome-length SARS-CoV cDNA template for in vitro transcription of SARS-CoV infectious RNA transcripts. Transfection of the RNA transcripts into permissive cells led to the recovery of infectious virus (recSARS-CoV). Characterization of the plaques produced by recSARS-CoV showed that they were similar in size to the parental SARS-CoV isolate HKU-39849 but smaller than the SARS-CoV isolate Frankfurt-1. Comparative analysis of replication kinetics showed that the kinetics of recSARS-CoV replication are similar to those of SARS-CoV Frankfurt-1, although the titers of virus released into the culture supernatant are approximately 10-fold less. The reverse genetic system was finally used to generate a recSARS-CoV reporter virus expressing Renilla luciferase in order to facilitate the analysis of SARS-CoV gene expression in human dendritic cells (hDCs). In parallel, a Renilla luciferase gene was also inserted into the genome of human coronavirus 229E (HCoV-229E). Using this approach, we demonstrate that, in contrast to HCoV-229E, SARS-CoV is not able to mediate efficient heterologous gene expression in hDCs.
DOI: 10.1371/journal.pone.0032857
PubMed: 22412934
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 001385
- to stream PubMed, to step Curation: 001385
- to stream PubMed, to step Checkpoint: 001301
- to stream Ncbi, to step Merge: 002475
- to stream Ncbi, to step Curation: 002475
- to stream Ncbi, to step Checkpoint: 002475
- to stream Main, to step Merge: 001C85
- to stream Main, to step Curation: 001C70
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Reverse genetics of SARS-related coronavirus using vaccinia virus-based recombination.</title>
<author><name sortKey="Van Den Worm, Sjoerd H E" sort="Van Den Worm, Sjoerd H E" uniqKey="Van Den Worm S" first="Sjoerd H E" last="Van Den Worm">Sjoerd H E. Van Den Worm</name>
<affiliation wicri:level="3"><nlm:affiliation>Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands.</nlm:affiliation>
<country xml:lang="fr">Pays-Bas</country>
<wicri:regionArea>Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden</wicri:regionArea>
<placeName><settlement type="city">Leyde</settlement>
<region nuts="2" type="province">Hollande-Méridionale</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Eriksson, Klara Kristin" sort="Eriksson, Klara Kristin" uniqKey="Eriksson K" first="Klara Kristin" last="Eriksson">Klara Kristin Eriksson</name>
</author>
<author><name sortKey="Zevenhoven, Jessika C" sort="Zevenhoven, Jessika C" uniqKey="Zevenhoven J" first="Jessika C" last="Zevenhoven">Jessika C. Zevenhoven</name>
</author>
<author><name sortKey="Weber, Friedemann" sort="Weber, Friedemann" uniqKey="Weber F" first="Friedemann" last="Weber">Friedemann Weber</name>
</author>
<author><name sortKey="Zust, Roland" sort="Zust, Roland" uniqKey="Zust R" first="Roland" last="Züst">Roland Züst</name>
</author>
<author><name sortKey="Kuri, Thomas" sort="Kuri, Thomas" uniqKey="Kuri T" first="Thomas" last="Kuri">Thomas Kuri</name>
</author>
<author><name sortKey="Dijkman, Ronald" sort="Dijkman, Ronald" uniqKey="Dijkman R" first="Ronald" last="Dijkman">Ronald Dijkman</name>
</author>
<author><name sortKey="Chang, Guohui" sort="Chang, Guohui" uniqKey="Chang G" first="Guohui" last="Chang">Guohui Chang</name>
</author>
<author><name sortKey="Siddell, Stuart G" sort="Siddell, Stuart G" uniqKey="Siddell S" first="Stuart G" last="Siddell">Stuart G. Siddell</name>
</author>
<author><name sortKey="Snijder, Eric J" sort="Snijder, Eric J" uniqKey="Snijder E" first="Eric J" last="Snijder">Eric J. Snijder</name>
</author>
<author><name sortKey="Thiel, Volker" sort="Thiel, Volker" uniqKey="Thiel V" first="Volker" last="Thiel">Volker Thiel</name>
</author>
<author><name sortKey="Davidson, Andrew D" sort="Davidson, Andrew D" uniqKey="Davidson A" first="Andrew D" last="Davidson">Andrew D. Davidson</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="2012">2012</date>
<idno type="RBID">pubmed:22412934</idno>
<idno type="pmid">22412934</idno>
<idno type="doi">10.1371/journal.pone.0032857</idno>
<idno type="wicri:Area/PubMed/Corpus">001385</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">001385</idno>
<idno type="wicri:Area/PubMed/Curation">001385</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">001385</idno>
<idno type="wicri:Area/PubMed/Checkpoint">001301</idno>
<idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">001301</idno>
<idno type="wicri:Area/Ncbi/Merge">002475</idno>
<idno type="wicri:Area/Ncbi/Curation">002475</idno>
<idno type="wicri:Area/Ncbi/Checkpoint">002475</idno>
<idno type="wicri:Area/Main/Merge">001C85</idno>
<idno type="wicri:Area/Main/Curation">001C70</idno>
<idno type="wicri:Area/Main/Exploration">001C70</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en">Reverse genetics of SARS-related coronavirus using vaccinia virus-based recombination.</title>
<author><name sortKey="Van Den Worm, Sjoerd H E" sort="Van Den Worm, Sjoerd H E" uniqKey="Van Den Worm S" first="Sjoerd H E" last="Van Den Worm">Sjoerd H E. Van Den Worm</name>
<affiliation wicri:level="3"><nlm:affiliation>Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden, The Netherlands.</nlm:affiliation>
<country xml:lang="fr">Pays-Bas</country>
<wicri:regionArea>Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, Leiden</wicri:regionArea>
<placeName><settlement type="city">Leyde</settlement>
<region nuts="2" type="province">Hollande-Méridionale</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Eriksson, Klara Kristin" sort="Eriksson, Klara Kristin" uniqKey="Eriksson K" first="Klara Kristin" last="Eriksson">Klara Kristin Eriksson</name>
</author>
<author><name sortKey="Zevenhoven, Jessika C" sort="Zevenhoven, Jessika C" uniqKey="Zevenhoven J" first="Jessika C" last="Zevenhoven">Jessika C. Zevenhoven</name>
</author>
<author><name sortKey="Weber, Friedemann" sort="Weber, Friedemann" uniqKey="Weber F" first="Friedemann" last="Weber">Friedemann Weber</name>
</author>
<author><name sortKey="Zust, Roland" sort="Zust, Roland" uniqKey="Zust R" first="Roland" last="Züst">Roland Züst</name>
</author>
<author><name sortKey="Kuri, Thomas" sort="Kuri, Thomas" uniqKey="Kuri T" first="Thomas" last="Kuri">Thomas Kuri</name>
</author>
<author><name sortKey="Dijkman, Ronald" sort="Dijkman, Ronald" uniqKey="Dijkman R" first="Ronald" last="Dijkman">Ronald Dijkman</name>
</author>
<author><name sortKey="Chang, Guohui" sort="Chang, Guohui" uniqKey="Chang G" first="Guohui" last="Chang">Guohui Chang</name>
</author>
<author><name sortKey="Siddell, Stuart G" sort="Siddell, Stuart G" uniqKey="Siddell S" first="Stuart G" last="Siddell">Stuart G. Siddell</name>
</author>
<author><name sortKey="Snijder, Eric J" sort="Snijder, Eric J" uniqKey="Snijder E" first="Eric J" last="Snijder">Eric J. Snijder</name>
</author>
<author><name sortKey="Thiel, Volker" sort="Thiel, Volker" uniqKey="Thiel V" first="Volker" last="Thiel">Volker Thiel</name>
</author>
<author><name sortKey="Davidson, Andrew D" sort="Davidson, Andrew D" uniqKey="Davidson A" first="Andrew D" last="Davidson">Andrew D. Davidson</name>
</author>
</analytic>
<series><title level="j">PloS one</title>
<idno type="eISSN">1932-6203</idno>
<imprint><date when="2012" type="published">2012</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Cell Line</term>
<term>Chlorocebus aethiops</term>
<term>Cloning, Molecular</term>
<term>DNA, Complementary</term>
<term>Dendritic Cells (virology)</term>
<term>Gene Expression Regulation, Viral</term>
<term>Gene Order</term>
<term>Genome, Viral</term>
<term>Humans</term>
<term>Molecular Sequence Data</term>
<term>Reassortant Viruses (genetics)</term>
<term>Recombination, Genetic</term>
<term>SARS Virus (genetics)</term>
<term>SARS Virus (growth & development)</term>
<term>Sequence Analysis, DNA</term>
<term>Vaccinia virus (genetics)</term>
<term>Viral Plaque Assay</term>
<term>Virus Replication</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ADN complémentaire</term>
<term>Analyse de séquence d'ADN</term>
<term>Animaux</term>
<term>Cellules dendritiques (virologie)</term>
<term>Clonage moléculaire</term>
<term>Données de séquences moléculaires</term>
<term>Génome viral</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Méthode des plages virales</term>
<term>Ordre des gènes</term>
<term>Recombinaison génétique</term>
<term>Régulation de l'expression des gènes viraux</term>
<term>Réplication virale</term>
<term>Virus de la vaccine (génétique)</term>
<term>Virus du SRAS (croissance et développement)</term>
<term>Virus du SRAS (génétique)</term>
<term>Virus recombinants (génétique)</term>
</keywords>
<keywords scheme="MESH" type="chemical" xml:lang="en"><term>DNA, Complementary</term>
</keywords>
<keywords scheme="MESH" qualifier="croissance et développement" xml:lang="fr"><term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Reassortant Viruses</term>
<term>SARS Virus</term>
<term>Vaccinia virus</term>
</keywords>
<keywords scheme="MESH" qualifier="growth & development" xml:lang="en"><term>SARS Virus</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Virus de la vaccine</term>
<term>Virus du SRAS</term>
<term>Virus recombinants</term>
</keywords>
<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Cellules dendritiques</term>
</keywords>
<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Dendritic Cells</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Cell Line</term>
<term>Chlorocebus aethiops</term>
<term>Cloning, Molecular</term>
<term>Gene Expression Regulation, Viral</term>
<term>Gene Order</term>
<term>Genome, Viral</term>
<term>Humans</term>
<term>Molecular Sequence Data</term>
<term>Recombination, Genetic</term>
<term>Sequence Analysis, DNA</term>
<term>Viral Plaque Assay</term>
<term>Virus Replication</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>ADN complémentaire</term>
<term>Analyse de séquence d'ADN</term>
<term>Animaux</term>
<term>Clonage moléculaire</term>
<term>Données de séquences moléculaires</term>
<term>Génome viral</term>
<term>Humains</term>
<term>Lignée cellulaire</term>
<term>Méthode des plages virales</term>
<term>Ordre des gènes</term>
<term>Recombinaison génétique</term>
<term>Régulation de l'expression des gènes viraux</term>
<term>Réplication virale</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) is a zoonotic disease caused by SARS-related coronavirus (SARS-CoV) that emerged in 2002 to become a global health concern. Although the original outbreak was controlled by classical public health measures, there is a real risk that another SARS-CoV could re-emerge from its natural reservoir, either in its original form or as a more virulent or pathogenic strain; in which case, the virus would be difficult to control in the absence of any effective antiviral drugs or vaccines. Using the well-studied SARS-CoV isolate HKU-39849, we developed a vaccinia virus-based SARS-CoV reverse genetic system that is both robust and biosafe. The SARS-CoV genome was cloned in separate vaccinia virus vectors, (vSARS-CoV-5prime and vSARS-CoV-3prime) as two cDNAs that were subsequently ligated to create a genome-length SARS-CoV cDNA template for in vitro transcription of SARS-CoV infectious RNA transcripts. Transfection of the RNA transcripts into permissive cells led to the recovery of infectious virus (recSARS-CoV). Characterization of the plaques produced by recSARS-CoV showed that they were similar in size to the parental SARS-CoV isolate HKU-39849 but smaller than the SARS-CoV isolate Frankfurt-1. Comparative analysis of replication kinetics showed that the kinetics of recSARS-CoV replication are similar to those of SARS-CoV Frankfurt-1, although the titers of virus released into the culture supernatant are approximately 10-fold less. The reverse genetic system was finally used to generate a recSARS-CoV reporter virus expressing Renilla luciferase in order to facilitate the analysis of SARS-CoV gene expression in human dendritic cells (hDCs). In parallel, a Renilla luciferase gene was also inserted into the genome of human coronavirus 229E (HCoV-229E). Using this approach, we demonstrate that, in contrast to HCoV-229E, SARS-CoV is not able to mediate efficient heterologous gene expression in hDCs.</div>
</front>
</TEI>
<affiliations><list><country><li>Pays-Bas</li>
</country>
<region><li>Hollande-Méridionale</li>
</region>
<settlement><li>Leyde</li>
</settlement>
</list>
<tree><noCountry><name sortKey="Chang, Guohui" sort="Chang, Guohui" uniqKey="Chang G" first="Guohui" last="Chang">Guohui Chang</name>
<name sortKey="Davidson, Andrew D" sort="Davidson, Andrew D" uniqKey="Davidson A" first="Andrew D" last="Davidson">Andrew D. Davidson</name>
<name sortKey="Dijkman, Ronald" sort="Dijkman, Ronald" uniqKey="Dijkman R" first="Ronald" last="Dijkman">Ronald Dijkman</name>
<name sortKey="Eriksson, Klara Kristin" sort="Eriksson, Klara Kristin" uniqKey="Eriksson K" first="Klara Kristin" last="Eriksson">Klara Kristin Eriksson</name>
<name sortKey="Kuri, Thomas" sort="Kuri, Thomas" uniqKey="Kuri T" first="Thomas" last="Kuri">Thomas Kuri</name>
<name sortKey="Siddell, Stuart G" sort="Siddell, Stuart G" uniqKey="Siddell S" first="Stuart G" last="Siddell">Stuart G. Siddell</name>
<name sortKey="Snijder, Eric J" sort="Snijder, Eric J" uniqKey="Snijder E" first="Eric J" last="Snijder">Eric J. Snijder</name>
<name sortKey="Thiel, Volker" sort="Thiel, Volker" uniqKey="Thiel V" first="Volker" last="Thiel">Volker Thiel</name>
<name sortKey="Weber, Friedemann" sort="Weber, Friedemann" uniqKey="Weber F" first="Friedemann" last="Weber">Friedemann Weber</name>
<name sortKey="Zevenhoven, Jessika C" sort="Zevenhoven, Jessika C" uniqKey="Zevenhoven J" first="Jessika C" last="Zevenhoven">Jessika C. Zevenhoven</name>
<name sortKey="Zust, Roland" sort="Zust, Roland" uniqKey="Zust R" first="Roland" last="Züst">Roland Züst</name>
</noCountry>
<country name="Pays-Bas"><region name="Hollande-Méridionale"><name sortKey="Van Den Worm, Sjoerd H E" sort="Van Den Worm, Sjoerd H E" uniqKey="Van Den Worm S" first="Sjoerd H E" last="Van Den Worm">Sjoerd H E. Van Den Worm</name>
</region>
</country>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/SrasV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001C70 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 001C70 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= SrasV1 |flux= Main |étape= Exploration |type= RBID |clé= pubmed:22412934 |texte= Reverse genetics of SARS-related coronavirus using vaccinia virus-based recombination. }}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:22412934" \ | HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \ | NlmPubMed2Wicri -a SrasV1
This area was generated with Dilib version V0.6.33. |