Examination of a plasmid-based reverse genetics system for human astrovirus

Benoit Chapellier, Shoichiro Tange, Hidetaka Tasaki, Kazuhiro Yoshida, Yan Zhou, Naomi Sakon, Kazuhiko Katayama, Akira Nakanishi

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)586-596
Number of pages11
JournalMICROBIOLOGY and IMMUNOLOGY
Volume59
Issue number10
DOIs
Publication statusPublished - Oct 2015
Externally publishedYes

Keywords

  • Animal RNA virus
  • Human astrovirus
  • Reverse genetics system

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Virology

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