1887

Abstract

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.

Funding
This study was supported by the:
  • Satoshi Komoto , Mochida Memorial Foundation for Medical and Pharmaceutical Research
  • Satoshi Komoto , Takeda Science Foundation
  • Tetsushi Yoshikawa , Japan Society for the Promotion of Science , (Award 18H02784)
  • Satoshi Komoto , Japan Society for the Promotion of Science , (Award 15K08505, 18K07150)
  • Satoshi Komoto , Japan Agency for Medical Research and Development , (Award 18fk0108018h0403, 18fk0108034h1102)
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2020-06-03
2020-09-22
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References

  1. Tate JE, Burton AH, Boschi-Pinto C, Parashar UD. World health Organization-Coordinated global rotavirus surveillance network. global, regional, and national estimates of rotavirus mortality in children. Clin Infect Dis 2016; 62:S96–S105
    [Google Scholar]
  2. Estes MK, Greenberg HB et al. Rotaviruses, p 1347-1401. In Knipe DM, Howley PM. (editors) Fields Virology, 6th ed. Philadelphia, PA: Lippincott Williams & Wilkins; 2013
    [Google Scholar]
  3. Desselberger U, Richards J, Tchertanov L, Lepault J, Lever A et al. Further characterisation of rotavirus cores: Ss(+)RNAs can be packaged in vitro but packaging lacks sequence specificity. Virus Res 2013; 178:252–263 [CrossRef][PubMed]
    [Google Scholar]
  4. Settembre EC, Chen JZ, Dormitzer PR, Grigorieff N, Harrison SC. Atomic model of an infectious rotavirus particle. Embo J 2011; 30:408–416 [CrossRef][PubMed]
    [Google Scholar]
  5. Viskovska M, Anish R, Hu L, Chow D-C, Hurwitz AM et al. Probing the sites of interactions of rotaviral proteins involved in replication. J Virol 2014; 88:12866–12881 [CrossRef][PubMed]
    [Google Scholar]
  6. Ramig RF. Genetics of the rotaviruses. Annu Rev Microbiol 1997; 51:225–255 [CrossRef][PubMed]
    [Google Scholar]
  7. Clark HF, Hoshino Y, Bell LM, Groff J, Hess G et al. Rotavirus isolate WI61 representing a presumptive new human serotype. J Clin Microbiol 1987; 25:1757–1762[PubMed]
    [Google Scholar]
  8. Greenberg HB, Kalica AR, Wyatt RG, Jones RW, Kapikian AZ et al. Rescue of noncultivatable human rotavirus by gene reassortment during mixed infection with Ts mutants of a cultivatable bovine rotavirus. Proc Natl Acad Sci U S A 1981; 78:420–424 [CrossRef][PubMed]
    [Google Scholar]
  9. Greenberg HB, Flores J, Kalica AR, Wyatt RG, Jones R. Gene coding assignments for growth restriction, neutralization and subgroup specificities of the W and DS-1 strains of human rotavirus. J Gen Virol 1983; 64 (Pt 2:313–320 [CrossRef][PubMed]
    [Google Scholar]
  10. Hoshino Y, Saif LJ, Sereno MM, Chanock RM, Kapikian AZ. Infection immunity of piglets to either VP3 or VP7 outer capsid protein confers resistance to challenge with a virulent rotavirus bearing the corresponding antigen. J Virol 1988; 62:744–748[PubMed]
    [Google Scholar]
  11. Hoshino Y, Saif LJ, Kang SY, Sereno MM, Chen WK et al. Identification of group A rotavirus genes associated with virulence of a porcine rotavirus and host range restriction of a human rotavirus in the gnotobiotic piglet model. Virology 1995; 209:274–280 [CrossRef][PubMed]
    [Google Scholar]
  12. Hoshino Y, Jones RW, Chanock RM, Kapikian AZ. Construction of four double gene substitution human X bovine rotavirus reassortant vaccine candidates: each bears two outer capsid human rotavirus genes, one encoding P serotype 1A and the other encoding G serotype 1, 2, 3, or 4 specificity. J Med Virol 1997; 51:319–325[PubMed]
    [Google Scholar]
  13. Kalica AR, Flores J, Greenberg HB. Identification of the rotaviral gene that codes for hemagglutination and protease-enhanced plaque formation. Virology 1983; 125:194–205 [CrossRef][PubMed]
    [Google Scholar]
  14. Kobayashi N, Taniguchi K, Urasawa T, Urasawa S. Efficient production of antigenic mosaic reassortants of rotavirus with the aid of anti-VP4 and anti-VP7 neutralizing monoclonal antibodies. J Virol Methods 1993; 44:25–34 [CrossRef][PubMed]
    [Google Scholar]
  15. Kobayashi N, Kojima K, Taniguchi K, Urasawa T, Urasawa S. Genotypic diversity of reassortants between simian rotavirus SA11 and human rotaviruses having different antigenic specificities and RNA patterns. Res Virol 1994; 145:303–311 [CrossRef][PubMed]
    [Google Scholar]
  16. Mahbub Alam M, Kobayashi N, Ishino M, Naik TN, Taniguchi K. Analysis of genetic factors related to preferential selection of the NSP1 gene segment observed in mixed infection and multiple passage of rotaviruses. Arch Virol 2006; 151:2149–2159 [CrossRef][PubMed]
    [Google Scholar]
  17. Midthun K, Greenberg HB, Hoshino Y, Kapikian AZ, Wyatt RG et al. Reassortant rotaviruses as potential live rotavirus vaccine candidates. J Virol 1985; 53:949–954[PubMed]
    [Google Scholar]
  18. Midthun K, Hoshino Y, Kapikian AZ, Chanock RM. Single gene substitution rotavirus reassortants containing the major neutralization protein (VP7) of human rotavirus serotype 4. J Clin Microbiol 1986; 24:822–826[PubMed]
    [Google Scholar]
  19. Okada J, Kobayashi N, Taniguchi K, Shiomi H. Functional analysis of the heterologous NSP1 genes in the genetic background of simian rotavirus SA11. Arch Virol 1999; 144:1439–1449 [CrossRef][PubMed]
    [Google Scholar]
  20. Okada J, Kobayashi N, Taniguchi K, Urasawa S. Analysis on reassortment of rotavirus NSP1 genes lacking coding region for cysteine-rich zinc finger motif. Arch Virol 1999; 144:345–353 [CrossRef][PubMed]
    [Google Scholar]
  21. Gombold JL, Ramig RF. Analysis of reassortment of genome segments in mice mixedly infected with rotaviruses SA11 and RRV. J Virol 1986; 57:110–116[PubMed]
    [Google Scholar]
  22. Ramig RF, Gombold JL. Rotavirus temperature-sensitive mutants are genetically stable and participate in reassortment during mixed infection of mice. Virology 1991; 182:468–474 [CrossRef][PubMed]
    [Google Scholar]
  23. Graham A, Kudesia G, Allen AM, Desselberger U. Reassortment of human rotavirus possessing genome rearrangements with bovine rotavirus: evidence for host cell selection. J Gen Virol 1987; 68 (Pt 1:115–122 [CrossRef][PubMed]
    [Google Scholar]
  24. Nakagomi O, Nakagomi T, Hoshino Y, Flores J, Kapikian AZ. Genetic analysis of a human rotavirus that belongs to subgroup I but has an RNA pattern typical of subgroup II human rotaviruses. J Clin Microbiol 1987; 25:1159–1164[PubMed]
    [Google Scholar]
  25. Nibert ML, Margraf RL, Coombs KM. Nonrandom segregation of parental alleles in reovirus reassortants. J Virol 1996; 70:7295–7300[PubMed]
    [Google Scholar]
  26. Ward RL, Nakagomi O, Knowlton DR, McNeal MM, Nakagomi T et al. Formation and selection of intergenogroup reassortants during cell culture adaptation of rotaviruses from dually infected subjects. J Virol 1991; 65:2699–2701[PubMed]
    [Google Scholar]
  27. Heiman EM, McDonald SM, Barro M, Taraporewala ZF, Bar-Magen T et al. Group a human rotavirus genomics: evidence that gene constellations are influenced by viral protein interactions. J Virol 2008; 82:11106–11116 [CrossRef][PubMed]
    [Google Scholar]
  28. McDonald SM, Matthijnssens J, McAllen JK, Hine E, Overton L et al. Evolutionary dynamics of human rotaviruses: balancing reassortment with preferred genome constellations. PLoS Pathog 2009; 5:e1000634 [CrossRef][PubMed]
    [Google Scholar]
  29. Johne R, Reetz J, Kaufer BB, Trojnar E. Generation of an avian-mammalian rotavirus reassortant by using a helper virus-dependent reverse genetics system. J Virol 2016; 90:1439–1443 [CrossRef][PubMed]
    [Google Scholar]
  30. Komoto S, Sasaki J, Taniguchi K. Reverse genetics system for introduction of site-specific mutations into the double-stranded RNA genome of infectious rotavirus. Proc Natl Acad Sci U S A 2006; 103:4646–4651 [CrossRef][PubMed]
    [Google Scholar]
  31. Navarro A, Trask SD, Patton JT. Generation of genetically stable recombinant rotaviruses containing novel genome rearrangements and heterologous sequences by reverse genetics. J Virol 2013; 87:6211–6220 [CrossRef][PubMed]
    [Google Scholar]
  32. Trask SD, Taraporewala ZF, Boehme KW, Dermody TS, Patton JT. Dual selection mechanisms drive efficient single-gene reverse genetics for rotavirus. Proc Natl Acad Sci U S A 2010; 107:18652–18657 [CrossRef][PubMed]
    [Google Scholar]
  33. Troupin C, Dehée A, Schnuriger A, Vende P, Poncet D et al. Rearranged genomic RNA segments offer a new approach to the reverse genetics of rotaviruses. J Virol 2010; 84:6711–6719 [CrossRef][PubMed]
    [Google Scholar]
  34. Kanai Y, Komoto S, Kawagishi T, Nouda R, Nagasawa N et al. Entirely plasmid-based reverse genetics system for rotaviruses. Proc Natl Acad Sci U S A 2017; 114:2349–2354 [CrossRef][PubMed]
    [Google Scholar]
  35. Komoto S, Kanai Y, Fukuda S, Kugita M, Kawagishi T et al. Reverse genetics system demonstrates that rotavirus nonstructural protein NSP6 is not essential for viral replication in cell culture. J Virol 2017; 91:e00695–17 [CrossRef][PubMed]
    [Google Scholar]
  36. Komoto S, Fukuda S, Ide T, Ito N, Sugiyama M et al. Generation of recombinant rotaviruses expressing fluorescent proteins by using an optimized reverse genetics system. J Virol 2018; 92:e00588–18 [CrossRef][PubMed]
    [Google Scholar]
  37. Komoto S, Fukuda S, Kugita M, Hatazawa R, Koyama C et al. Generation of infectious recombinant human rotaviruses from just 11 cloned cDNAs encoding the rotavirus genome. J Virol 2019; 93:e02207–02218 [CrossRef][PubMed]
    [Google Scholar]
  38. Falkenhagen A, Patzina-Mehling C, Rückner A, Vahlenkamp TW, Johne R. Generation of simian rotavirus reassortants with diverse VP4 genes using reverse genetics. J Gen Virol 2019; 100:1595–1604 [CrossRef][PubMed]
    [Google Scholar]
  39. Taniguchi K, Nishikawa K, Kobayashi N, Urasawa T, Wu H et al. Differences in plaque size and VP4 sequence found in SA11 virus clones having simian authentic VP4. Virology 1994; 198:325–330 [CrossRef][PubMed]
    [Google Scholar]
  40. Falkenhagen A, Patzina-Mehling C, Gadicherla AK, Strydom A, O'Neill HG et al. Generation of simian rotavirus reassortants with VP4- and VP7-encoding genome segments from human strains circulating in Africa using reverse genetics. Viruses 2020; 12:E201 [CrossRef][PubMed]
    [Google Scholar]
  41. Matthijnssens J, Heylen E, Zeller M, Rahman M, Lemey P et al. Phylodynamic analyses of rotavirus genotypes G9 and G12 underscore their potential for swift global spread. Mol Biol Evol 2010; 27:2431–2436 [CrossRef][PubMed]
    [Google Scholar]
  42. O'Ryan M, O’Ryan M. The ever-changing landscape of rotavirus serotypes. Pediatr Infect Dis J 2009; 28:S60–S62 [CrossRef][PubMed]
    [Google Scholar]
  43. Burke RM, Tate JE, Kirkwood CD, Steele AD, Parashar UD. Current and new rotavirus vaccines. Curr Opin Infect Dis 2019; 32:435–444 [CrossRef][PubMed]
    [Google Scholar]
  44. Soares-Weiser K, Bergman H, Henschke N, Pitan F, Cunliffe N. Vaccines for preventing rotavirus diarrhoea: vaccines in use. Cochrane Database Syst Rev 2019; 2019: [CrossRef][PubMed]
    [Google Scholar]
  45. Bowen MD, Mijatovic-Rustempasic S, Esona MD, Teel EN, Gautam R et al. Rotavirus strain trends during the postlicensure vaccine era: United States, 2008-2013. J Infect Dis 2016; 214:732–738 [CrossRef][PubMed]
    [Google Scholar]
  46. Ogden KM, Tan Y, Akopov A, Stewart LS, McHenry R et al. Multiple introductions and antigenic mismatch with vaccines may contribute to increased predominance of G12P[8] rotaviruses in the United States. J Virol 2019; 93:e01476–18 [CrossRef][PubMed]
    [Google Scholar]
  47. Kawagishi T, Nurdin JA, Onishi M, Nouda R, Kanai Y et al. Reverse genetics system for a human group A rotavirus. J Virol 2020; 94:e00963–19 [CrossRef][PubMed]
    [Google Scholar]
  48. Ito N, Takayama-Ito M, Yamada K, Hosokawa J, Sugiyama M et al. Improved recovery of rabies virus from cloned cDNA using a vaccinia virus-free reverse genetics system. Microbiol Immunol 2003; 47:613–617 [CrossRef][PubMed]
    [Google Scholar]
  49. Urasawa S, Urasawa T, Taniguchi K, Chiba S. Serotype determination of human rotavirus isolates and antibody prevalence in pediatric population in Hokkaido, Japan. Arch Virol 1984; 81:1–12 [CrossRef][PubMed]
    [Google Scholar]
  50. Green KY, Sears JF, Taniguchi K, Midthun K, Hoshino Y et al. Prediction of human rotavirus serotype by nucleotide sequence analysis of the VP7 protein gene. J Virol 1988; 62:1819–1823[PubMed]
    [Google Scholar]
  51. Clark HF, Offit PA, Ellis RW, Eiden JJ, Krah D et al. The development of multivalent bovine rotavirus (strain WC3) reassortant vaccine for infants. J Infect Dis 1996; 174 Suppl 1:S73–S80 [CrossRef][PubMed]
    [Google Scholar]
  52. Taniguchi K, Urasawa T, Kobayashi N, Gorziglia M, Urasawa S. Nucleotide sequence of VP4 and VP7 genes of human rotaviruses with subgroup I specificity and long RNA pattern: implication for new G serotype specificity. J Virol 1990; 64:5640–5644[PubMed]
    [Google Scholar]
  53. Schnell MJ, Mebatsion T, Conzelmann KK. Infectious rabies viruses from cloned cDNA. Embo J 1994; 13:4195–4203[PubMed]
    [Google Scholar]
  54. Taniguchi K, Morita Y, Urasawa T, Urasawa S. Cross-Reactive neutralization epitopes on VP3 of human rotavirus: analysis with monoclonal antibodies and antigenic variants. J Virol 1987; 61:1726–1730[PubMed]
    [Google Scholar]
  55. Urasawa S, Urasawa T, Taniguchi K. Three human rotavirus serotypes demonstrated by plaque neutralization of isolated strains. Infect Immun 1982; 38:781–784[PubMed]
    [Google Scholar]
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