TY - JOUR
T1 - Examination of a plasmid-based reverse genetics system for human astrovirus
AU - Chapellier, Benoit
AU - Tange, Shoichiro
AU - Tasaki, Hidetaka
AU - Yoshida, Kazuhiro
AU - Zhou, Yan
AU - Sakon, Naomi
AU - Katayama, Kazuhiko
AU - Nakanishi, Akira
N1 - Publisher Copyright:
© 2015 The Societies and Wiley Publishing Asia Pty Ltd.
PY - 2015/10
Y1 - 2015/10
N2 - 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.
AB - 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.
KW - Animal RNA virus
KW - Human astrovirus
KW - Reverse genetics system
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U2 - 10.1111/1348-0421.12317
DO - 10.1111/1348-0421.12317
M3 - Article
C2 - 26272702
AN - SCOPUS:84944456529
VL - 59
SP - 586
EP - 596
JO - Microbiology and Immunology
JF - Microbiology and Immunology
SN - 0385-5600
IS - 10
ER -