Small interfering RNA targeting m2 gene induces effective and long term inhibition of influenza A virus replication.
RNA interference (RNAi) provides a powerful new means to inhibit viral infection specifically. However, the selection of siRNA-resistant viruses is a major concern in the use of RNAi as antiviral therapeutics. In this study, we conducted a lentiviral vector with a H1-short hairpin RNA (shRNA) expression cassette to deliver small interfering RNAs (siRNAs) into mammalian cells. Using this vector that also expresses enhanced green fluorescence protein (EGFP) as surrogate marker, stable shRNA-expressing cell lines were successfully established and the inhibition efficiencies of rationally designed siRNAs targeting to conserved regions of influenza A virus genome were assessed. The results showed that a siRNA targeting influenza M2 gene (siM2) potently inhibited viral replication. The siM2 was not only effective for H1N1 virus but also for highly pathogenic avian influenza virus H5N1. In addition to its M2 inhibition, the siM2 also inhibited NP mRNA accumulation and protein expression. A long term inhibition effect of the siM2 was demonstrated and the emergence of siRNA-resistant mutants in influenza quasispecies was not observed. Taken together, our study suggested that M2 gene might be an optimal RNAi target for antiviral therapy. These findings provide useful information for the development of RNAi-based prophylaxis and therapy for human influenza virus infection.
Sui
Hong-Yan
HY
Department of Microbiology, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.
Zhao
Guang-Yu
GY
Huang
Jian-Dong
JD
Jin
Dong-Yan
DY
Yuen
Kwok-Yung
KY
Zheng
Bo-Jian
BJ
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004285:Dogs; D015967:Gene Expression Regulation, Viral; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000502:physiology*; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism; D009976:Orthomyxoviridae Infections / Q000821:virology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D012692:Serial Passage; D013997:Time Factors; D014763:Viral Matrix Proteins / Q000235:genetics*; D019065:Virus Assembly; D014779:Virus Replication / Q000502:physiology*; D000086082:Viroporin Proteins
D019590:Nucleocapsid Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins