Efficient genome engineering of Toxoplasma gondii using the TALEN technique
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 mark...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-03-01
|
| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13071-019-3378-y |
| _version_ | 1819261780588756992 |
|---|---|
| author | Hongmei Chen Yijia Guo Yushu Qiu Huanbin Huang Changqing Lin Min Liu Xiaoguang Chen Peiliang Yang Kun Wu |
| author_facet | Hongmei Chen Yijia Guo Yushu Qiu Huanbin Huang Changqing Lin Min Liu Xiaoguang Chen Peiliang Yang Kun Wu |
| author_sort | Hongmei Chen |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-23T19:47:14Z |
| format | Article |
| id | doaj.art-3d503258e8fb49948da6a40e86c3008b |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-12-23T19:47:14Z |
| publishDate | 2019-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-3d503258e8fb49948da6a40e86c3008b2022-12-21T17:33:29ZengBMCParasites & Vectors1756-33052019-03-0112111010.1186/s13071-019-3378-yEfficient genome engineering of Toxoplasma gondii using the TALEN techniqueHongmei Chen0Yijia Guo1Yushu Qiu2Huanbin Huang3Changqing Lin4Min Liu5Xiaoguang Chen6Peiliang Yang7Kun Wu8Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityExperimental Animal Center, Nanfang Hospital, Southern Medical UniversityDepartment of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical UniversityAbstract 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.http://link.springer.com/article/10.1186/s13071-019-3378-yTALENToxoplasma gondiiBradyzoiteTyrosine hydroxylase 2In vivo imaging |
| spellingShingle | Hongmei Chen Yijia Guo Yushu Qiu Huanbin Huang Changqing Lin Min Liu Xiaoguang Chen Peiliang Yang Kun Wu Efficient genome engineering of Toxoplasma gondii using the TALEN technique Parasites & Vectors TALEN Toxoplasma gondii Bradyzoite Tyrosine hydroxylase 2 In vivo imaging |
| title | Efficient genome engineering of Toxoplasma gondii using the TALEN technique |
| title_full | Efficient genome engineering of Toxoplasma gondii using the TALEN technique |
| title_fullStr | Efficient genome engineering of Toxoplasma gondii using the TALEN technique |
| title_full_unstemmed | Efficient genome engineering of Toxoplasma gondii using the TALEN technique |
| title_short | Efficient genome engineering of Toxoplasma gondii using the TALEN technique |
| title_sort | efficient genome engineering of toxoplasma gondii using the talen technique |
| topic | TALEN Toxoplasma gondii Bradyzoite Tyrosine hydroxylase 2 In vivo imaging |
| url | http://link.springer.com/article/10.1186/s13071-019-3378-y |
| work_keys_str_mv | AT hongmeichen efficientgenomeengineeringoftoxoplasmagondiiusingthetalentechnique AT yijiaguo efficientgenomeengineeringoftoxoplasmagondiiusingthetalentechnique AT yushuqiu efficientgenomeengineeringoftoxoplasmagondiiusingthetalentechnique AT huanbinhuang efficientgenomeengineeringoftoxoplasmagondiiusingthetalentechnique AT changqinglin efficientgenomeengineeringoftoxoplasmagondiiusingthetalentechnique AT minliu efficientgenomeengineeringoftoxoplasmagondiiusingthetalentechnique AT xiaoguangchen efficientgenomeengineeringoftoxoplasmagondiiusingthetalentechnique AT peiliangyang efficientgenomeengineeringoftoxoplasmagondiiusingthetalentechnique AT kunwu efficientgenomeengineeringoftoxoplasmagondiiusingthetalentechnique |