Small interfering RNA inhibits SARS-CoV nucleocapsid gene expression in cultured cells and mouse muscles.
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.
Zhao
Ping
P
Department of Microbiology, Second Military Medical University, Shanghai, China.
Qin
Zhao-Ling
ZL
Ke
Jin-Shan
JS
Lu
Yang
Y
Liu
Min
M
Pan
Wei
W
Zhao
Lan-Juan
LJ
Cao
Jie
J
Qi
Zhong-Tian
ZT
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*
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