A cell-based screening system for influenza A viral RNA transcription/replication inhibitors.
Although two classes of antivirals, NA inhibitors and M2 ion channel blockers, are licensed for influenza treatment, dual resistant mutants, including highly pathogenic H5N1 viruses, have appeared. Alternative treatment options are, therefore, needed. Influenza A viral RNA (vRNA) transcription/replication is a promising target for antiviral development, since it is essential for virus replication. Accordingly, an efficient and reliable method to identify vRNA transcription/replication inhibitors is desirable. Here, we developed a cell-based screening system by establishing a cell line that stably expresses influenza viral ribonucleoprotein complex (vRNP). Compound library screening using this cell line allowed us to identify a compound that inhibits vRNA transcription/replication by using reporter protein expression from virus-like RNA as a readout and virus replication in vitro. vRNP-expressing cells have potential as a simple and convenient high-throughput screening (HTS) system, and, thus, are promising to identify vRNA transcription/replication inhibitors for various RNA viruses, especially for primary screens.
Department of Special Pathogens, International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
Makoto
M
Ozawa
Masayuki
M
Shimojima
Hideo
H
Goto
Shinji
S
Watanabe
Yasuko
Y
Hatta
Maki
M
Kiso
Yousuke
Y
Furuta
Taisuke
T
Horimoto
Noel R
NR
Peters
F Michael
FM
Hoffmann
Yoshihiro
Y
Kawaoka
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D004285:Dogs; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009980:Influenza A virus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology*; D007251:Influenza, Human / Q000188:drug therapy / Q000821:virology; D061985:Madin Darby Canine Kidney Cells; D012328:RNA Viruses / Q000187:drug effects / Q000235:genetics; D012367:RNA, Viral / Q000187:drug effects* / Q000235:genetics; D020394:Vault Ribonucleoprotein Particles / Q000187:drug effects / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects* / Q000235:genetics
D000998:Antiviral Agents; D012367:RNA, Viral; D020394:Vault Ribonucleoprotein Particles; D014764:Viral Proteins