CRISPR-Cas9-based method for isolating microgametes of Eimeria tenella

文献类型： 外文期刊

第一作者： Qu, Zigang

作者： Qu, Zigang;Olajide, Joshua Seun;Wang, Jing;Cai, Jianping;Gong, Zhenxing;Qu, Zigang;Cai, Jianping;Olajide, Joshua Seun

作者机构：

关键词： Eimeria tenella; GCS1 gene; Microgamete; CRISPR-Cas9; Genome editing; Flow cytometry

期刊名称：VETERINARY PARASITOLOGY （ 影响因子：2.6； 五年影响因子：2.5 ）

ISSN： 0304-4017

年卷期： 2024 年 327 卷

页码：

收录情况： SCI

摘要： Eimeria tenella infections are known to cause severe caecal damage and death of the infected chicken. Gamogony is an essential stage in E. tenella life cycle and in the establishment of coccidiosis. Prior research had extensively explored isolation and separation of the parasite gametes - microgamete (male) and macrogamete (female). However, there is little information on the efficient, highly purified and distinctly separated male and female gametes. In this study, we generated a genome editing line expressing mCherry fluorescent protein fused with GCS1 protein in E. tenella by using Toxoplasma gondii CRISPR-Cas9 system, flow cytometry and fluorescence microscopy. This allowed precise separation of E. tenella male and female gametes in the transgenic parasite population. The separation of male and female gametes would not only build on our understanding of E. tenella transmission, but it would also facilitate development of gametocidal compounds as drug targets for E. tenella infection.

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