Development of an optimized RNA-based murine norovirus reverse genetics system.

Journal of Virological Methods
Muhammad A YunusIan G. Goodfellow

Abstract

Murine norovirus (MNV), identified in 2003, is the only norovirus which replicates efficiently in tissue culture and as a result has been used extensively as a model for human noroviruses, a major cause of acute gastroenteritis. The current report describes the generation of a new approach to reverse genetics recovery of genetically defined MNV that relies on the transfection of in vitro transcribed capped RNA directly into cells. The use of the recently developed ScriptCap post-transcriptional enzymatic capping system, followed by optimized Neon mediated electroporation of the highly permissive RAW 264.7 cells, resulted in the rapid and robust recovery of infectious MNV. Transfection of cells capable of supporting virus replication but not permissive to virus infection, namely human or hamster kidney cells, also resulted in robust recovery of infectious virus without subsequent amplification by multiple rounds of re-infection. This latter system may provide a reproducible method to measure the specific infectivity of mutant norovirus RNA allowing the accurate quantitation of the effect of mutations on norovirus replication.

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Citations

Aug 31, 2012·Journal of Virology·Surender VashistIan Goodfellow
Aug 24, 2012·Journal of Virology·Lucy ThorneIan Goodfellow
Jul 5, 2012·Journal of Visualized Experiments : JoVE·Armando AriasIan Goodfellow
Sep 7, 2014·Proceedings of the National Academy of Sciences of the United States of America·Kazuhiko KatayamaMary K Estes
Dec 15, 2015·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Yu LiuYun-Xing Wang
Aug 15, 2015·Microbiology and Immunology·Benoit ChapellierAkira Nakanishi
Oct 12, 2014·The Journal of General Virology·Myra HosmilloKyoung-Oh Cho
Nov 19, 2013·The Journal of General Virology·Lucy G Thorne, Ian G Goodfellow
Sep 4, 2012·Journal of Virological Methods·Maël Bessaud, Francis Delpeyroux
Dec 3, 2014·Trends in Microbiology·Stefan Taube, Christiane E Wobus
Feb 24, 2015·Journal of Virological Methods·Carlos Sandoval-JaimeStanislav V Sosnovtsev
Jul 16, 2014·Journal of Virological Methods·Tomoichiro OkaYukinobu Tohya
Mar 14, 2018·Antimicrobial Agents and Chemotherapy·Mitsutaka KitanoIan Goodfellow
Feb 17, 2018·Archives of Virology·Yifeng QinWeijian Huang
Aug 16, 2013·Journal of Virology·Eduardo López-ManríquezAna Lorena Gutiérrez-Escolano
Feb 7, 2014·Journal of Virology·Abimbola O KolawoleChristiane E Wobus
Apr 5, 2015·Applied and Environmental Microbiology·Tun-Yun Hsueh, Kristen E Gibson
Apr 10, 2015·Journal of Virology·Surender VashistIan Goodfellow
Jun 4, 2019·Viruses·Vincent R GrazianoCraig B Wilen
Feb 6, 2020·Pharmaceutics·Abishek WadhwaAneesh Thakur
Jul 3, 2020·PLoS Pathogens·Chihong SongKazuyoshi Murata
Jan 1, 2017·Uirusu·Kazuhiko Katayama
Apr 4, 2020·The Journal of General Virology·Louisa F Ludwig-BegallEtienne Thiry
Dec 26, 2018·Medicinal Research Reviews·Natalie E NetzlerPeter A White
Feb 15, 2019·Viruses·Kyle V Todd, Ralph A Tripp
Mar 25, 2019·Viruses·Ulrich Desselberger
May 3, 2014·Current Protocols in Microbiology·Seungmin HwangChristiane E Wobus
Dec 12, 2020·Frontiers in Microbiology·Ángel L ÁlvarezFrancisco Parra
Jun 23, 2021·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Longfei LiuYingwei Wang
Aug 29, 2021·Viruses·Louisa F Ludwig-BegallEtienne Thiry

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Methods Mentioned

BETA
transfection
PCR
in vitro transcription
FCS
fluorescence
flow cytometry

Software Mentioned

ScriptCap
Epicentre

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