New reverse genetics and transfection methods to rescue arboviruses in mosquito cells View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2017-12

AUTHORS

Thérèse Atieh, Antoine Nougairède, Raphaëlle Klitting, Fabien Aubry, Anna-Bella Failloux, Xavier de Lamballerie, Stéphane Priet

ABSTRACT

Reverse genetics is a critical tool to decrypt the biological properties of arboviruses. However, whilst reverse genetics methods have been usually applied to vertebrate cells, their use in insect cells remains uncommon due to the conjunction of laborious molecular biology techniques and of specific difficulties surrounding the transfection of such cells. To leverage reverse genetics studies in both vertebrate and mosquito cells, we designed an improved DNA transfection protocol for insect cells and then demonstrated that the simple and flexible ISA (Infectious Subgenomic Amplicons) reverse-genetics method can be efficiently applied to both mammalian and mosquito cells to generate in days recombinant infectious positive-stranded RNA viruses belonging to genera Flavivirus (Japanese encephalitis, Yellow fever, West Nile and Zika viruses) and Alphavirus (Chikungunya virus). This method represents an effective option to potentially overcome technological issues related to the study of arboviruses. More... »

PAGES

13983

Journal

TITLE

Scientific Reports

ISSUE

1

VOLUME

7

From Grant

  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/s41598-017-14522-6

    DOI

    http://dx.doi.org/10.1038/s41598-017-14522-6

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/29070887


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