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Parasit Vectors. 2019 Mar 15;12(1):112. doi: 10.1186/s13071-019-3378-y.

Efficient genome engineering of Toxoplasma gondii using the TALEN technique.

Author information

1
Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China.
2
Experimental Animal Center, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China. plyang@smu.edu.cn.
3
Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China. wksmu@163.com.

Abstract

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.

KEYWORDS:

Bradyzoite; In vivo imaging; TALEN; Toxoplasma gondii; Tyrosine hydroxylase 2

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