Suppression of the Sendai virus M protein through a novel short interfering RNA approach inhibits viral particle production but does not affect viral RNA synthesis.
Short RNA interference is more and more widely recognized as an effective method to specifically suppress viral functions in eukaryotic cells. Here, we used an experimental system that allows suppression of the Sendai virus (SeV) M protein by using a target sequence, derived from the green fluorescent protein gene, that was introduced in the 3' untranslated region of the M protein mRNA. Silencing of the M protein gene was eventually achieved by a small interfering RNA (siRNA) directed against this target sequence. This siRNA was constitutively expressed in a cell line constructed by transduction with an appropriate lentivirus vector. Suppression of the M protein was sufficient to diminish virus production by 50- to 100-fold. This level of suppression had no apparent effect on viral replication and transcription, supporting the lack of M involvement in SeV transcription or replication control.
Mottet-Osman
Geneviève
G
Department of Microbiology and Molecular Medicine, University of Geneva Medical School, CMU, 1 Rue Michel-Servet, CH-1211 Geneva 4, Switzerland. laurent.roux@medecine.unige.ch
Iseni
Frédéric
F
Pelet
Thierry
T
Wiznerowicz
Maciej
M
Garcin
Dominique
D
Roux
Laurent
L
D045744:Cell Line, Tumor; D020868:Gene Silencing; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism*; D012367:RNA, Viral / Q000096:biosynthesis*; D014161:Transduction, Genetic; D014162:Transfection; D014763:Viral Matrix Proteins / Q000235:genetics*; D014771:Virion / Q000378:metabolism*
C097020:M protein, Sendai virus; D034741:RNA, Small Interfering; D012367:RNA, Viral; D014763:Viral Matrix Proteins; D049452:Green Fluorescent Proteins