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Efficient genome engineering of Toxoplasma gondii using the TALEN technique
- Chen, Hongmei, Guo, Yijia, Qiu, Yushu, Huang, Huanbin, Lin, Changqing, Liu, Min, Chen, Xiaoguang, Yang, Peiliang, Wu, Kun
- Parasites & vectors 2019 v.12 no.1 pp. 112
- Toxoplasma gondii, amino acids, animal models, enzymes, fluorescence, genetic engineering, genetically modified organisms, green fluorescent protein, host-parasite relationships, image analysis, parasites, pyrimethamine, screening
- BACKGROUND: Aromatic amino acid hydroxylase 2 (AAH2) is a bradyzoite-specific upregulated protein that may alter host behaviour by altering the host dopaminergic pathway. To better understand the role of the parasite’s AAH2 in host-parasite interactions, we generated an AAH2 fluorescent marker strain of T. gondii using the TALEN technique. METHODS: We generated an AAH2 fluorescent marker strain of T. gondii, which was designated PRU/AAH2-eGFP, using the TALEN technique. This strain stably expressed pyrimethamine resistance for screening and expressed enhanced green fluorescent protein (eGFP)-tagged AAH2 in the bradyzoite stage. The bradyzoite conversion of PRU/AAH2-eGFP was observed both in vitro and in vivo. The fluorescence localization of AAH2 in mouse models of chronic infection was observed by a Bruker in vivo imaging system. RESULTS: Transgenic T. gondii was successfully generated by the TALEN system. The eGFP-tagged AAH2 could be detected by in vivo imaging. CONCLUSIONS: This study verified the feasibility of using TALEN technology for T. gondii research and provided an in vivo imaging method for in vivo research of bradyzoite-stage proteins.