Dual infections by group A and group C rotaviruses have been reported, but no reassortants between group A and group C rotaviruses have been described. The VP6 major inner capsid protein of group A and C rotaviruses shares common antigens detected by monoclonal antibodies and also shares 40-43% amino acid identity. Coinfection of Spodoptera frugiperda (Sf9) insect cells with different combinations of the recombinant baculoviruses encoding either group A [RF VP2 (A-VP2), IND VP6 (A-VP6), and VP7 (A-VP7[IND]), 2292B VP7 (A-VP7[2292B])] or C [Shintoku VP6 (C-VP6) and VP7 (C-VP7)] bovine rotavirus proteins produced hybrid group A/C triple-layered VP2/6/7 virus-like particles (TLPs) composed of A-VP2/C-VP6/C-VP7, A-VP2/C-VP6/A-VP7(IND), A-VP2/C-VP6/A-VP7(2292B), and A-VP2/A-VP6/C-VP7. To our knowledge, this is the first report that the inner capsid VP6 of group A or group C rotavirus can support attachment of the heterologous, antigenically distinct group A (G6, IND or G10, 2292B) or group C rotavirus outer capsid VP7 to produce hybrid TLPs in vitro.