Fig. 6

Infectivity potential of NcGRA11 (a–e). a Strategy for NcGRA11 (a–e) disruption by CRISPR/Cas9-mediated homologous gene replacement. b Diagnostic PCRs on two ∆ncgra11 (a–e) clones (1 and 2). (F4-R4) and (F5-R5) check the correct integration of the selection marker to the NcGRA11 (a–e) genes locus, whereas (F6-R6) examines the deletion of the NcGRA11 (a–e) genes. c Western blotting with NcGRA11c anti-mouse antibodies with NcGRA11c of 91 kDa in the parent Nc1 strain. No NcGRA11c protein was detected in the ∆ncgra11 (a–e) polyclonal strains (1, 2, 3, and 4). NcActin was used as a loading control. d Plaque assay comparing the growth of Nc1 and ∆ncgra11 (a–e). Each well was infected with 200 parasites, and plaques were stained 9 days later. e The plaque areas were counted by randomly selecting at least 20 plaques and using the pixel point in Photoshop C6S software (Adobe, USA). The data were compiled from two independent experiments. Statistical analysis was performed as above. f Mouse survival after infection with 5 × 10⁶ or 8 × 10⁶ doses of ∆ncgra11 (a–e) and Nc1. There were five female mice in each group. Statistical analysis was performed as above