RISPR/CAS9技术在构建弓形虫微线体蛋白MIC3敲除株上的应用

he recently discovered CRISPR/CAS9 technique has become an important genome editing tool in the molecular biology and cell biology research fields, and played critical roles in studies on host infection by pathogens. Toxoplasma gondii is an obligate intracellular protozoan, which can infects almost all warm-blooded animals. Micronemal proteins are secreted by micronemes, which are unique organelles involved in host infection by Toxoplasma gondii. To further understand the functions of micronemal proteins in host infection, we focused on the MIC3 gene and made genetic knock-outs in RH and PRU strains using the CRISPR/CAS9 system. After positive clone screening and identification, the knockout strains have been successful constructed. This study demonstrated that CRISPR/CAS9 could be used for gene knock-outs in a variety of Toxoplasma gondii strains with different genetic backgrounds.