Rotavirus gene silencing by small interfering RNAs.
RNA interference is an evolutionarily conserved double-stranded RNA-triggered mechanism for suppressing gene expression. Rotaviruses, the leading cause of severe diarrhea in young children, are formed by three concentric layers of protein, from which the spike protein VP4 projects. Here, we show that a small interfering RNA corresponding to the VP4 gene efficiently inhibits the synthesis of this protein in virus-infected cells. A large proportion of infected cells had no detectable VP4 and the yield of viral progeny was reduced. Most of the virus particles purified from these cells were triple-layered, but lacked VP4, and were poorly infectious. We also show that VP4 might not be required for the last step of virus morphogenesis. The VP4 gene silencing was specific, since the synthesis of VP4 from rotavirus strains that differ in the target sequence was not affected. These findings offer the possibility of carrying out reverse genetics in rotaviruses.
Déctor
Miguel Angel
MA
Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Cuernavaca, Morelos 62210, Mexico.
Romero
Pedro
P
López
Susana
S
Arias
Carlos F
CF
D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D020868:Gene Silencing; D005814:Genes, Viral; D034622:RNA Interference; D034741:RNA, Small Interfering; D012401:Rotavirus / Q000235:genetics* / Q000254:growth & development
D036022:Capsid Proteins; D034741:RNA, Small Interfering; C060339:VP4 protein, Rotavirus