Examination of a plasmid-based reverse genetics system for human astrovirus.
A plasmid-based reverse genetics system for human astrovirus type 1 (HAstV1) is examined. Upon transfection into 293T cells, the plasmid vector, which harbors a HAstV1 expression cassette, expressed astroviral RNA that appeared to be capable of viral RNA replication, as indicated by the production of subgenomic RNA and capsid protein expression irrespective of the heterologous 5' ends of the transcribed RNA. Particles infectious to Caco-2 cells were made in this system; however, their infectivity was much lower than would be expected from the amount of particles apparently produced. Using Huh-7 cells as the transfection host with the aim of improving viral capsid processing for virion maturation partially restored the efficiency of infectious particle formation. Our results support the possibility that the DNA transfection process induces a cellular response that targets late, but not early, stages of HAstV1 infection.
Laboratory of Gene Therapy, Department of Aging Intervention, National Center for Geriatrics and Gerontology, Obu, Aichi, 474-8522.
Benoit
B
Chapellier
Laboratory of Gene Therapy, Department of Aging Intervention, National Center for Geriatrics and Gerontology, Obu, Aichi, 474-8522.
Shoichiro
S
Tange
Laboratory of Gene Therapy, Department of Aging Intervention, National Center for Geriatrics and Gerontology, Obu, Aichi, 474-8522.
Hidetaka
H
Tasaki
Laboratory of Gene Therapy, Department of Aging Intervention, National Center for Geriatrics and Gerontology, Obu, Aichi, 474-8522.
Kazuhiro
K
Yoshida
Laboratory of Gene Therapy, Department of Aging Intervention, National Center for Geriatrics and Gerontology, Obu, Aichi, 474-8522.
Yan
Y
Zhou
Osaka Prefectural Institute of Public Health, Osaka, Osaka, 537-0025.
Naomi
N
Sakon
National Institute for Infectious Diseases, Department of Virology 2, Laboratory of Gastroenteritis Viruses, Musashi-Murayama, Tokyo, 208-0011, Japan.
Kazuhiko
K
Katayama
Laboratory of Gene Therapy, Department of Aging Intervention, National Center for Geriatrics and Gerontology, Obu, Aichi, 474-8522.
Akira
A
Nakanishi
D019350:Astroviridae Infections / Q000235:genetics / Q000821:virology; D018938:Caco-2 Cells; D036022:Capsid Proteins / Q000096:biosynthesis*; D002460:Cell Line; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000276:immunology; D005759:Gastroenteritis / Q000821:virology; D057809:HEK293 Cells; D006801:Humans; D007113:Immunity, Innate / Q000276:immunology; D001257:Mamastrovirus / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics*; D011971:Receptors, Immunologic; D059386:Reverse Genetics; D014162:Transfection; D019065:Virus Assembly / Q000235:genetics; D014779:Virus Replication / Q000235:genetics*
D036022:Capsid Proteins; D012367:RNA, Viral; D011971:Receptors, Immunologic; C452820:RIGI protein, human; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases
animal RNA virus; human astrovirus; reverse genetics system