Inhibition of SARS-CoV replication cycle by small interference RNAs silencing specific sars proteins, 7a/7b, 3a/3b and s
Identifieur interne : 003B58 ( Main/Exploration ); précédent : 003B57; suivant : 003B59Inhibition of SARS-CoV replication cycle by small interference RNAs silencing specific sars proteins, 7a/7b, 3a/3b and s
Auteurs : Sara Akerström [Suède] ; Ali Mirazimi [Suède] ; Yee-Joo Tan [Singapour]Source :
- Antiviral research [ 0166-3542 ] ; 2007.
Descripteurs français
- KwdFr :
- ARN messager (génétique), Animaux, Cellules Vero, Petit ARN interférent (génétique), Protéines de la matrice virale (biosynthèse), Protéines de la matrice virale (génétique), Protéines virales (biosynthèse), Protéines virales (génétique), Réplication virale (génétique), Spécificité du substrat, Syndrome respiratoire aigu sévère (), Syndrome respiratoire aigu sévère (génétique), Syndrome respiratoire aigu sévère (virologie), Technique d'immunofluorescence (), Virus du SRAS (génétique), Virus du SRAS (physiologie).
- MESH :
- biosynthèse : Protéines de la matrice virale, Protéines virales.
- génétique : ARN messager, Petit ARN interférent, Protéines de la matrice virale, Protéines virales, Réplication virale, Syndrome respiratoire aigu sévère, Virus du SRAS.
- physiologie : Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- Pascal (Inist)
English descriptors
- KwdEn :
- Animals, Antiviral, Chlorocebus aethiops, Coronavirus, Fluorescent Antibody Technique (methods), Gene silencing, Protein, RNA interference, RNA, Messenger (genetics), RNA, Small Interfering (genetics), Replication, SARS Virus (genetics), SARS Virus (physiology), Severe Acute Respiratory Syndrome (genetics), Severe Acute Respiratory Syndrome (therapy), Severe Acute Respiratory Syndrome (virology), Severe acute respiratory syndrome, Severe acute respiratory syndrome virus, Small Interference RNA, Substrate Specificity, Vero Cells, Viral Matrix Proteins (biosynthesis), Viral Matrix Proteins (genetics), Viral Proteins (biosynthesis), Viral Proteins (genetics), Virus Replication (genetics).
- MESH :
- chemical , biosynthesis : Viral Matrix Proteins, Viral Proteins.
- chemical , genetics : RNA, Messenger, RNA, Small Interfering, Viral Matrix Proteins, Viral Proteins.
- genetics : SARS Virus, Severe Acute Respiratory Syndrome, Virus Replication.
- methods : Fluorescent Antibody Technique.
- physiology : SARS Virus.
- therapy : Severe Acute Respiratory Syndrome.
- virology : Severe Acute Respiratory Syndrome.
- Animals, Chlorocebus aethiops, Substrate Specificity, Vero Cells.
Abstract
The severe acute respiratory syndrome coronavirus (SARS CoV) genome has 14 potential open reading frames (ORFs). The first ORF is translated from the full-length genomic mRNA while the remaining ORFs are translated from eight subgeomic RNAs (sgRNAs). In this study, we designed small interference RNAs (siRNAs) targeting sgRNA 2, 3 and 7 and tested their efficiency and specificity in silencing the protein translated from the targeted sgRNA. Our results demonstrated that siRNA 7 could inhibit sgRNA 7, which showed 19/19 nucleotides (nt) matching, and sgRNA 8, which showed 18/19 nt matching; but, it did not inhibit the full-length genomic mRNA which showed 17/19 nt matching. Overall, each of the siRNAs can inhibit the targeted sgRNA without affecting the full-length genomic mRNA or the other sgRNAs that showed mismatch of two or more nt. Thus, siRNA could be designed so as to knockdown the expression of viral protein(s) from a targeted sgRNA during viral infection, thereby allowing the contribution of individual viral proteins to viral infection to be delineated. When Vero E6 cells expressing siRNA 2, 3 or 7 were infected with SARS-CoV, a significant reduction in the yield of progeny virus was observed. Indirect immunofluorescence assays showed that in the infected cells expressing each of the siRNAs, there was aspecific silencing of S, 3a and 7a, respectively, but the expression of nucleocapsid protein was not affected. Thus, our data suggests that the accessory proteins, i.e. 3a and 7a, could play an important role during the replication cycle of the SARS-CoV.
Affiliations:
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Le document en format XML
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for Infectious Disease Control</s1><s2>171 82 Solna</s2><s3>SWE</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ></inist:fA14><country>Suède</country><wicri:noRegion>171 82 Solna</wicri:noRegion></affiliation></author><author><name sortKey="Tan, Yee Joo" sort="Tan, Yee Joo" uniqKey="Tan Y" first="Yee-Joo" last="Tan">Yee-Joo Tan</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Institute of Molecular and Cell Biology, 61 Biopolis Drive</s1><s2>138673 Singapore</s2><s3>SGP</s3><sZ>3 aut.</sZ></inist:fA14><country>Singapour</country><wicri:noRegion>138673 Singapore</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Antiviral research</title><title level="j" type="abbreviated">Antivir. res.</title><idno type="ISSN">0166-3542</idno><imprint><date when="2007">2007</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Antiviral research</title><title level="j" type="abbreviated">Antivir. res.</title><idno 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(biosynthesis)</term><term>Viral Proteins (genetics)</term><term>Virus Replication (genetics)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN messager (génétique)</term><term>Animaux</term><term>Cellules Vero</term><term>Petit ARN interférent (génétique)</term><term>Protéines de la matrice virale (biosynthèse)</term><term>Protéines de la matrice virale (génétique)</term><term>Protéines virales (biosynthèse)</term><term>Protéines virales (génétique)</term><term>Réplication virale (génétique)</term><term>Spécificité du substrat</term><term>Syndrome respiratoire aigu sévère ()</term><term>Syndrome respiratoire aigu sévère (génétique)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Technique d'immunofluorescence ()</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="biosynthesis" xml:lang="en"><term>Viral Matrix Proteins</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Messenger</term><term>RNA, Small Interfering</term><term>Viral Matrix Proteins</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" qualifier="biosynthèse" xml:lang="fr"><term>Protéines de la matrice virale</term><term>Protéines virales</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term><term>Severe Acute Respiratory Syndrome</term><term>Virus Replication</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN messager</term><term>Petit ARN interférent</term><term>Protéines de la matrice virale</term><term>Protéines virales</term><term>Réplication virale</term><term>Syndrome respiratoire aigu sévère</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Fluorescent Antibody Technique</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="therapy" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Substrate Specificity</term><term>Vero Cells</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>Coronavirus</term><term>Spécificité du substrat</term><term>Syndrome respiratoire aigu sévère</term><term>Réplication</term><term>Technique d'immunofluorescence</term><term>siRNA</term><term>Protéine</term><term>Silence expression génique</term><term>Virus syndrome respiratoire aigu sévère</term><term>Antiviral</term><term>Interférence ARN</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The severe acute respiratory syndrome coronavirus (SARS CoV) genome has 14 potential open reading frames (ORFs). The first ORF is translated from the full-length genomic mRNA while the remaining ORFs are translated from eight subgeomic RNAs (sgRNAs). In this study, we designed small interference RNAs (siRNAs) targeting sgRNA 2, 3 and 7 and tested their efficiency and specificity in silencing the protein translated from the targeted sgRNA. Our results demonstrated that siRNA 7 could inhibit sgRNA 7, which showed 19/19 nucleotides (nt) matching, and sgRNA 8, which showed 18/19 nt matching; but, it did not inhibit the full-length genomic mRNA which showed 17/19 nt matching. Overall, each of the siRNAs can inhibit the targeted sgRNA without affecting the full-length genomic mRNA or the other sgRNAs that showed mismatch of two or more nt. Thus, siRNA could be designed so as to knockdown the expression of viral protein(s) from a targeted sgRNA during viral infection, thereby allowing the contribution of individual viral proteins to viral infection to be delineated. When Vero E6 cells expressing siRNA 2, 3 or 7 were infected with SARS-CoV, a significant reduction in the yield of progeny virus was observed. Indirect immunofluorescence assays showed that in the infected cells expressing each of the siRNAs, there was aspecific silencing of S, 3a and 7a, respectively, but the expression of nucleocapsid protein was not affected. Thus, our data suggests that the accessory proteins, i.e. 3a and 7a, could play an important role during the replication cycle of the SARS-CoV.</div></front></TEI><affiliations><list><country><li>Singapour</li><li>Suède</li></country></list><tree><country name="Suède"><noRegion><name sortKey="Akerstrom, Sara" sort="Akerstrom, Sara" uniqKey="Akerstrom S" first="Sara" last="Akerström">Sara Akerström</name></noRegion><name sortKey="Mirazimi, Ali" sort="Mirazimi, Ali" uniqKey="Mirazimi A" first="Ali" last="Mirazimi">Ali Mirazimi</name></country><country name="Singapour"><noRegion><name sortKey="Tan, Yee Joo" sort="Tan, Yee Joo" uniqKey="Tan Y" first="Yee-Joo" last="Tan">Yee-Joo Tan</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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