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© Christine Schmitt, Anubis Vega Rua, Jean-Marc Panaud
Tête de moustique femelle Aedes albopictus, vecteur du virus de la dengue et du chikungunya. Microphotographie électronique à balayage, image colorisée.
Publication : Scientific reports

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

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Scientific reports - 25 Oct 2017

Atieh T, Nougairède A, Klitting R, Aubry F, Failloux AB, de Lamballerie X, Priet S

Link to Pubmed [PMID] – 29070887

Sci Rep 2017 Oct;7(1):13983

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.

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