Deleting ku80 improves the efficiency of targeted gene editing in Neospora caninum

CRISPR/Cas9 technology has been widely used for gene editing in organisms. Gene deletion of the ku80/ku70 complex can improve the efficiency of gene replacement in Arabidopsis thaliana, Cryptococcus neoformans, and Toxoplasma gondii, which remained elusive in Neospora caninum. Here, we knock out the ku80 gene in Nc1 strain by using CRISPR/Cas9, detect the growth rate and virulence of Nc delta ku80. Then we compare the efficiency of gene replacements between Nc delta ku80 and Nc1 strains by transfected with the same HA-tagged plasmids, and the percentage of HA-tagged parasites was investigated by IFA. The results showed that gene targeting efficiency was increased in the Nc delta ku80 strain via double crossover at several genetic loci, but its growth rate and virulence were unaffected. In conclusion, the Nc delta ku80 strain can be used as an effective strain for rapid gene editing of N. caninum.

Automated summary

This study successfully knocked out the ku80 gene in the Nc1 strain using CRISPR/Cas9 technology, and demonstrated that the Nc delta ku80 strain can improve gene replacement efficiency without affecting the growth rate and virulence. It can be used as an effective strain for rapid gene editing of N. caninum.