Inhibition of genes expression of SARS coronavirus by synthetic small interfering RNAs.
RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA), and results in the silencing of homologous gene expression through the specific degradation of an mRNA containing the same sequence. dsRNA-mediated RNAi can be used in a wide variety of eucaryotes to induce the sequence-specific inhibition of gene expression. Synthetic 21-23 nucleotide (nt) small interfering RNA (siRNA) with 2 nt 3' overhangs was recently found to mediate efficient sequence-specific mRNA degradation in mammalian cells. Here, we studied the effects of synthetic siRNA duplexes targeted to SARS coronavirus structural proteins E, M, and N in a cell culture system. Among total 26 siRNA duplexes, we obtained 3 siRNA duplexes which could sequence-specifically reduce target genes expression over 80% at the concentration of 60 nM in Vero E6 cells. The downregulation effect was in correlation with the concentrations of the siRNA duplexes in a range of 0 approximately 60 nM. Our results also showed that many inactive siRNA duplexes may be brought to life simply by unpairing the 5'end of the antisense strands. Results suggest that siRNA is capable of inhibiting SARS coronavirus genes expression and thus may be a new therapeutic strategy for treatment of SARS.
Shi
Yi
Y
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Yang
De Hua
DH
Xiong
Jie
J
Jia
Jie
J
Huang
Bing
B
Jin
You Xin
YX
D000818:Animals; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D000086862:Coronavirus M Proteins; D000086462:Coronavirus Nucleocapsid Proteins; D015967:Gene Expression Regulation, Viral; D049452:Green Fluorescent Proteins / Q000378:metabolism; D019590:Nucleocapsid Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology; D045473:Severe acute respiratory syndrome-related coronavirus / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D014763:Viral Matrix Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D015678:Viral Structural Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D000086082:Viroporin Proteins
D000086862:Coronavirus M Proteins; D000086462:Coronavirus Nucleocapsid Proteins; C501689:E protein, SARS coronavirus; C000706237:M protein, SARS-CoV; D019590:Nucleocapsid Proteins; D034741:RNA, Small Interfering; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; D015678:Viral Structural Proteins; D000086082:Viroporin Proteins; D049452:Green Fluorescent Proteins