Main content area

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.