title: Small interfering RNA inhibits SARS-CoV nucleocapsid gene expression in cultured cells and mouse muscles.
abstract: SARS-CoV is a newly identified coronavirus that causes severe acute respiratory syndrome (SARS). Currently, there is no effective method available for prophylaxis and treatment of SARS-CoV infections. In the present study, the influence of small interfering RNA (siRNA) on SARS-CoV nucleocapsid (N) protein expression was detected in cultured cells and mouse muscles. Four siRNA expression cassettes driven by mouse U6 promoter targeting SARS-CoV N gene were prepared, and their inhibitory effects on expression of N and enhanced green fluorescence protein (EGFP) fusion protein were observed. A candidate siRNA was proved to down-regulate N and EGFP expression actively in a sequence-specific manner. The expression vector of this siRNA was constructed and confirmed to reduce N and EGFP expression efficiently in both cultured cells and adult mouse muscles. Our findings suggest that the siRNA should provide the basis for prophylaxis and therapy of SARS-CoV infection in human.
authors.affiliation: Department of Microbiology, Second Military Medical University, Shanghai, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhao
authors.forename: Zhao-Ling
authors.initials: ZL
authors.lastname: Qin
authors.forename: Jin-Shan
authors.initials: JS
authors.lastname: Ke
authors.forename: Yang
authors.initials: Y
authors.lastname: Lu
authors.forename: Min
authors.initials: M
authors.lastname: Liu
authors.forename: Wei
authors.initials: W
authors.lastname: Pan
authors.forename: Lan-Juan
authors.initials: LJ
authors.lastname: Zhao
authors.forename: Jie
authors.initials: J
authors.lastname: Cao
authors.forename: Zhong-Tian
authors.initials: ZT
authors.lastname: Qi
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D015870:Gene Expression / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D009132:Muscles / Q000378:metabolism*; D019590:Nucleocapsid Proteins / Q000235:genetics* / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000378:metabolism*
chemical_list: D019590:Nucleocapsid Proteins; D034741:RNA, Small Interfering; D012342:RNA, Small Nuclear; D011994:Recombinant Proteins; C405486:U6 small nuclear RNA; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins