In vitro culture and genetic modification of Babesia gibsoni
Babesia gibsoni, a unicellular eukaryotic parasite causing babesiosis in dog, is primarily transmitted through tick feeding. The intraerythrocytic stage, during which the parasite reproduce within the host's red blood cells, is a vital part of Babesia's life cycle. Continuous in vitro cult...
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| Format: | Article |
| Language: | English |
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KeAi Communications Co., Ltd.
2024-01-01
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| Series: | Decoding Infection and Transmission |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S294992402400003X |
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| author | Dongfang Li Sen Wang Xingai Guan Yidan Bai Junlong Zhao Lan He |
| author_facet | Dongfang Li Sen Wang Xingai Guan Yidan Bai Junlong Zhao Lan He |
| author_sort | Dongfang Li |
| collection | DOAJ |
| description | Babesia gibsoni, a unicellular eukaryotic parasite causing babesiosis in dog, is primarily transmitted through tick feeding. The intraerythrocytic stage, during which the parasite reproduce within the host's red blood cells, is a vital part of Babesia's life cycle. Continuous in vitro culture B. gibsoni provides an opportunity to study its biological processes. The establishment and development of gene editing systems for Babesia offer a powerful tool to investigate the functions of important genes in specific biological processes. This protocol expands on the existing techniques for in vitro culture and genes editing of B. gibsoni. Specifically, we describe a continuous in vitro culture method employing VP-SFM as a base medium, supplemented with Albumax I and small amount of canine serum (2.5 %), This method, designed for long-term culture, achieving high parasitemia and facilitates subclone culture. By employing homology-dependent repair pathways, the gene editing method utilizing introducing homologous fragments and electroporation can effectively manipulate the genetic of B. gibsoni. This protocol would contribute to the reproducibility of experiments and the overall reliability of research findings. |
| format | Article |
| id | doaj-art-bc29d0ed5861483dbbd6f4be60c29fc6 |
| institution | OA Journals |
| issn | 2949-9240 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Decoding Infection and Transmission |
| spelling | doaj-art-bc29d0ed5861483dbbd6f4be60c29fc62025-08-20T02:33:43ZengKeAi Communications Co., Ltd.Decoding Infection and Transmission2949-92402024-01-01210001910.1016/j.dcit.2024.100019In vitro culture and genetic modification of Babesia gibsoniDongfang Li0Sen Wang1Xingai Guan2Yidan Bai3Junlong Zhao4Lan He5State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Wuhan, 430070, Hubei, ChinaState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Wuhan, 430070, Hubei, ChinaState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Wuhan, 430070, Hubei, ChinaState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Wuhan, 430070, Hubei, ChinaState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, China; Key Laboratory of Animal Epidemical Disease and Infectious Zoonoses, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, Hubei, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Wuhan, 430070, Hubei, China; Corresponding author. State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, China; Key Laboratory of Animal Epidemical Disease and Infectious Zoonoses, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, Hubei, China; Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Wuhan, 430070, Hubei, China; Corresponding author. State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, Hubei, China.Babesia gibsoni, a unicellular eukaryotic parasite causing babesiosis in dog, is primarily transmitted through tick feeding. The intraerythrocytic stage, during which the parasite reproduce within the host's red blood cells, is a vital part of Babesia's life cycle. Continuous in vitro culture B. gibsoni provides an opportunity to study its biological processes. The establishment and development of gene editing systems for Babesia offer a powerful tool to investigate the functions of important genes in specific biological processes. This protocol expands on the existing techniques for in vitro culture and genes editing of B. gibsoni. Specifically, we describe a continuous in vitro culture method employing VP-SFM as a base medium, supplemented with Albumax I and small amount of canine serum (2.5 %), This method, designed for long-term culture, achieving high parasitemia and facilitates subclone culture. By employing homology-dependent repair pathways, the gene editing method utilizing introducing homologous fragments and electroporation can effectively manipulate the genetic of B. gibsoni. This protocol would contribute to the reproducibility of experiments and the overall reliability of research findings.http://www.sciencedirect.com/science/article/pii/S294992402400003XBabesia gibsoniErythrocyteIn vitro cultureGenetic editingHomology-dependent repair |
| spellingShingle | Dongfang Li Sen Wang Xingai Guan Yidan Bai Junlong Zhao Lan He In vitro culture and genetic modification of Babesia gibsoni Decoding Infection and Transmission Babesia gibsoni Erythrocyte In vitro culture Genetic editing Homology-dependent repair |
| title | In vitro culture and genetic modification of Babesia gibsoni |
| title_full | In vitro culture and genetic modification of Babesia gibsoni |
| title_fullStr | In vitro culture and genetic modification of Babesia gibsoni |
| title_full_unstemmed | In vitro culture and genetic modification of Babesia gibsoni |
| title_short | In vitro culture and genetic modification of Babesia gibsoni |
| title_sort | in vitro culture and genetic modification of babesia gibsoni |
| topic | Babesia gibsoni Erythrocyte In vitro culture Genetic editing Homology-dependent repair |
| url | http://www.sciencedirect.com/science/article/pii/S294992402400003X |
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