Involvement of INS15 in the development and pathogenicity of the zoonotic pathogen Cryptosporidium parvum
Fig 3
INS15 and INS16 are not essential for parasite survival.
(A-B) Schematic shows the knockout strategy of INS15 and INS16. The top line shows the native locus of INS15 or INS16, the middle line shows the repair template, and the bottom line shows the modified locus. sgRNA, single guide RNA; Nluc, nano-luciferase; NeoR, neomycin resistance marker; Eno, enolase promoter. P1, P2, P3, and P4 are primers used to verify genomic integration events. (C-D) Diagnosis PCR of the Δins15 and Δins16 locus. Fecal genomic DNA from WT, Δins15 and Δins16 parasites were used as template, and primers for checking 5’ insertion (5’ Ins), 3’ insertion (3’ Ins), and the 3’ UTR sequence of INS16 as a positive control (+ve) are indicated. (E-F) Growth patterns of Δins15 and INS15-HA strain (C) or Δins16 and INS16-HA strains (D) in HCT-8 cell cultures. Data from three independent experiments, bars are standard deviations; p = exact values shown as determined with two-way ANOVA corrected for multiple comparisons according to Sidak’s method.