Efficient genome engineering of Toxoplasma gondii using the TALEN technique

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

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.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Cells, Cultured
  • Feasibility Studies
  • Female
  • Fluorescent Dyes
  • Genetic Engineering / methods*
  • Genetic Markers
  • Genome, Protozoan*
  • Green Fluorescent Proteins / genetics
  • Host-Parasite Interactions
  • Humans
  • Mice
  • Microscopy, Fluorescence
  • Protozoan Proteins / genetics*
  • Toxoplasma / genetics*
  • Toxoplasmosis, Animal / parasitology
  • Transcription Activator-Like Effector Nucleases / genetics*

Substances

  • Fluorescent Dyes
  • Genetic Markers
  • Protozoan Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Transcription Activator-Like Effector Nucleases