TY - JOUR
T1 - Rapid generation of rotavirus single-gene reassortants by means of eleven plasmid-only based reverse genetics
AU - Fukuda, Saori
AU - Hatazawa, Riona
AU - Kawamura, Yoshiki
AU - Yoshikawa, Tetsushi
AU - Murata, Takayuki
AU - Taniguchi, Koki
AU - Komoto, Satoshi
N1 - Funding Information:
This study was supported in part by the MEXT-Supported Programme for the Research Programme on Emerging and Re-emerging Infectious Diseases of the Japan Agency for Medical Research and Development, AMED (18fk0108018h0403 and 18fk0108034h1102) (S. K.), JSPS KAKENHI (15K08505) (S. K.), 18K07150 (S. K.) and 18H02784 (T. Y.), the Takeda Science Foundation (S. K.) and The Mochida Memorial Foundation for Medical and Pharmaceutical Research (S. K.).
PY - 2020
Y1 - 2020
N2 - Reassortment is an important mechanism in the evolution of group A rotaviruses (RVAs), yielding viruses with novel genetic and phenotypic traits. The classical methods for generating RVA reassortants with the desired genetic combinations are laborious and time-consuming because of the screening and selection processes required to isolate a desired reassortant. Taking advantage of a recently developed RVA reverse genetics system based on just 11 cloned cDNAs encoding the RVA genome (11 plasmid-only system), we prepared a panel of simian SA11-L2 virus-based single-gene reassortants, each containing 1 segment derived from human KU virus of the G1P[8] genotype. It was shown that there was no gene-specific restriction of the reassortment potential. In addition to these 11 single-gene reassortants, a triple-gene reassortant with KU-derived core-encoding VP1–3 gene segments with the SA11-L2 genetic background, which make up a virion composed of the KU-based core, and SA11-L2-based intermediate and outer layers, could also be prepared with the 11 plasmid-only system. Finally, for possible clinical application of this system, we generated a series of VP7 reassortants representing all the major human RVA G genotypes (G1–4, G9 and G12) efficiently. The preparation of each of these single-gene reassortants was achieved within just 2 weeks. Our results demonstrate that the 11 plasmid-only system allows the rapid and reliable generation of RVA single-gene reassortants, which will be useful for basic research and clinical applications.
AB - Reassortment is an important mechanism in the evolution of group A rotaviruses (RVAs), yielding viruses with novel genetic and phenotypic traits. The classical methods for generating RVA reassortants with the desired genetic combinations are laborious and time-consuming because of the screening and selection processes required to isolate a desired reassortant. Taking advantage of a recently developed RVA reverse genetics system based on just 11 cloned cDNAs encoding the RVA genome (11 plasmid-only system), we prepared a panel of simian SA11-L2 virus-based single-gene reassortants, each containing 1 segment derived from human KU virus of the G1P[8] genotype. It was shown that there was no gene-specific restriction of the reassortment potential. In addition to these 11 single-gene reassortants, a triple-gene reassortant with KU-derived core-encoding VP1–3 gene segments with the SA11-L2 genetic background, which make up a virion composed of the KU-based core, and SA11-L2-based intermediate and outer layers, could also be prepared with the 11 plasmid-only system. Finally, for possible clinical application of this system, we generated a series of VP7 reassortants representing all the major human RVA G genotypes (G1–4, G9 and G12) efficiently. The preparation of each of these single-gene reassortants was achieved within just 2 weeks. Our results demonstrate that the 11 plasmid-only system allows the rapid and reliable generation of RVA single-gene reassortants, which will be useful for basic research and clinical applications.
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U2 - 10.1099/jgv.0.001443
DO - 10.1099/jgv.0.001443
M3 - Article
C2 - 32490794
AN - SCOPUS:85086938338
VL - 101
SP - 806
EP - 815
JO - Journal of General Virology
JF - Journal of General Virology
SN - 0022-1317
IS - 8
ER -