SrasV1, Main, Exploration, bibRecord, 005065

Silencing of SARS-CoV spike gene by small interfering RNA in HEK 293T cells.

Identifieur interne : 005065 ( Main/Exploration ); précédent : 005064; suivant : 005066

Silencing of SARS-CoV spike gene by small interfering RNA in HEK 293T cells.

Auteurs : Zhao-Ling Qin [République populaire de Chine] ; Ping Zhao ; Xiao-Lian Zhang ; Jian-Guo Yu ; Ming-Mei Cao ; Lan-Juan Zhao ; Jie Luan ; Zhong-Tian Qi

Source :

Biochemical and biophysical research communications [ 0006-291X ] ; 2004.

RBID : pubmed:15504339

Descripteurs français

KwdFr :
- Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (biosynthèse), Glycoprotéines membranaires (génétique), Humains, Interférence par ARN, Lignée cellulaire, Petit ARN interférent (biosynthèse), Petit ARN interférent (génétique), Petit ARN interférent (pharmacologie), Protéines de l'enveloppe virale (biosynthèse), Protéines de l'enveloppe virale (génétique), Virus du SRAS (génétique).
MESH :
- biosynthèse : Glycoprotéines membranaires, Petit ARN interférent, Protéines de l'enveloppe virale.
- génétique : Glycoprotéines membranaires, Petit ARN interférent, Protéines de l'enveloppe virale, Virus du SRAS.
- pharmacologie : Petit ARN interférent.
- Glycoprotéine de spicule des coronavirus, Humains, Interférence par ARN, Lignée cellulaire.

English descriptors

KwdEn :
- Cell Line, Humans, Membrane Glycoproteins (biosynthesis), Membrane Glycoproteins (genetics), RNA Interference, RNA, Small Interfering (biosynthesis), RNA, Small Interfering (genetics), RNA, Small Interfering (pharmacology), SARS Virus (genetics), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (biosynthesis), Viral Envelope Proteins (genetics).
MESH :
- chemical , biosynthesis : Membrane Glycoproteins, RNA, Small Interfering, Viral Envelope Proteins.
- chemical , genetics : Membrane Glycoproteins, RNA, Small Interfering, Viral Envelope Proteins.
- chemical , pharmacology : RNA, Small Interfering.
- genetics : SARS Virus.
- Cell Line, Humans, RNA Interference, Spike Glycoprotein, Coronavirus.

Abstract

Two candidate small interfering RNAs (siRNAs) corresponding to severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike gene were designed and in vitro transcribed to explore the possibility of silencing SARS-CoV S gene. The plasmid pEGFP-optS, which contains the codon-optimized SARS-CoV S gene and expresses spike-EGFP fusion protein (S-EGFP) as silencing target and expressing reporter, was transfected with siRNAs into HEK 293T cells. At various time points of posttransfection, the levels of S-EGFP expression and amounts of spike mRNA transcript were detected by fluorescence microscopy, flow cytometry, Western blot, and real-time quantitative PCR, respectively. The results showed that the cells transfected with pEGFP-optS expressed S-EGFP fusion protein at a higher level compared with those transfected with pEGFP-S, which contains wildtype SARS-CoV spike gene sequence. The green fluorescence, mean fluorescence intensity, and SARS-CoV S RNA transcripts were found significantly reduced, and the expression of SARS-CoV S glycoprotein was strongly inhibited in those cells co-transfected with either EGFP- or S-specific siRNAs. Our findings demonstrated that the S-specific siRNAs used in this study were able to specifically and effectively inhibit SARS-CoV S glycoprotein expression in cultured cells through blocking the accumulation of S mRNA, which may provide an approach for studies on the functions of SARS-CoV S gene and development of novel prophylactic or therapeutic agents for SARS-CoV.

DOI: 10.1016/j.bbrc.2004.09.180
PubMed: 15504339

Affiliations:

République populaire de Chine

Le document en format XML

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<author><name sortKey="Luan, Jie" sort="Luan, Jie" uniqKey="Luan J" first="Jie" last="Luan">Jie Luan</name>
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<author><name sortKey="Qi, Zhong Tian" sort="Qi, Zhong Tian" uniqKey="Qi Z" first="Zhong-Tian" last="Qi">Zhong-Tian Qi</name>
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<term>RNA Interference</term>
<term>RNA, Small Interfering (biosynthesis)</term>
<term>RNA, Small Interfering (genetics)</term>
<term>RNA, Small Interfering (pharmacology)</term>
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<term>Spike Glycoprotein, Coronavirus</term>
<term>Viral Envelope Proteins (biosynthesis)</term>
<term>Viral Envelope Proteins (genetics)</term>
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<term>Petit ARN interférent (génétique)</term>
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<term>Protéines de l'enveloppe virale (génétique)</term>
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<term>Viral Envelope Proteins</term>
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<front><div type="abstract" xml:lang="en">Two candidate small interfering RNAs (siRNAs) corresponding to severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike gene were designed and in vitro transcribed to explore the possibility of silencing SARS-CoV S gene. The plasmid pEGFP-optS, which contains the codon-optimized SARS-CoV S gene and expresses spike-EGFP fusion protein (S-EGFP) as silencing target and expressing reporter, was transfected with siRNAs into HEK 293T cells. At various time points of posttransfection, the levels of S-EGFP expression and amounts of spike mRNA transcript were detected by fluorescence microscopy, flow cytometry, Western blot, and real-time quantitative PCR, respectively. The results showed that the cells transfected with pEGFP-optS expressed S-EGFP fusion protein at a higher level compared with those transfected with pEGFP-S, which contains wildtype SARS-CoV spike gene sequence. The green fluorescence, mean fluorescence intensity, and SARS-CoV S RNA transcripts were found significantly reduced, and the expression of SARS-CoV S glycoprotein was strongly inhibited in those cells co-transfected with either EGFP- or S-specific siRNAs. Our findings demonstrated that the S-specific siRNAs used in this study were able to specifically and effectively inhibit SARS-CoV S glycoprotein expression in cultured cells through blocking the accumulation of S mRNA, which may provide an approach for studies on the functions of SARS-CoV S gene and development of novel prophylactic or therapeutic agents for SARS-CoV.</div>
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<name sortKey="Qi, Zhong Tian" sort="Qi, Zhong Tian" uniqKey="Qi Z" first="Zhong-Tian" last="Qi">Zhong-Tian Qi</name>
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Silencing of SARS-CoV spike gene by small interfering RNA in HEK 293T cells.

Silencing of SARS-CoV spike gene by small interfering RNA in HEK 293T cells.

Source :

Descripteurs français

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri

Pour générer des pages wiki

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