de Fabritus L., Nougairede A., Aubry F., Gould E. A., de Lamballerie Xavier. (2016). Utilisation of ISA reverse genetics and large-scale random codon re-encoding to produce attenuated strains of tick-borne encephalitis virus within days. Plos One, 11 (8), p. e0159564 [11 p.]. ISSN 1932-6203.
Titre du document
Utilisation of ISA reverse genetics and large-scale random codon re-encoding to produce attenuated strains of tick-borne encephalitis virus within days
Large-scale codon re-encoding is a new method of attenuating RNA viruses. However, the use of infectious clones to generate attenuated viruses has inherent technical problems. We previously developed a bacterium-free reverse genetics protocol, designated ISA, and now combined it with large-scale random codon-re-encoding method to produce attenuated tick-borne encephalitis virus (TBEV), a pathogenic flavivirus which causes febrile illness and encephalitis in humans. We produced wild-type (WT) and two re-encoded TBEVs, containing 273 or 273+284 synonymous mutations in the NS5 and NS5+NS3 coding regions respectively. Both re-encoded viruses were attenuated when compared with WT virus using a laboratory mouse model and the relative level of attenuation increased with the degree of re-encoding. Moreover, all infected animals produced neutralizing antibodies. This novel, rapid and efficient approach to engineering attenuated viruses could potentially expedite the development of safe and effective new-generation live attenuated vaccines.
