Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system
Abstract Eimeria tenella has emerged as valuable model organism for studying the biology and immunology of protozoan parasites with the establishment of the reverse genetic manipulation platform. In this report, we described the application of CRISPR (clustered regularly interspaced short palindromi...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2020-03-01
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| Series: | Veterinary Research |
| Online Access: | http://link.springer.com/article/10.1186/s13567-020-00766-0 |
| _version_ | 1818908415080005632 |
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| author | Xinming Tang Jingxia Suo Lin Liang Chunhui Duan Dandan Hu Xiaolong Gu Yonglan Yu Xianyong Liu Shangjin Cui Xun Suo |
| author_facet | Xinming Tang Jingxia Suo Lin Liang Chunhui Duan Dandan Hu Xiaolong Gu Yonglan Yu Xianyong Liu Shangjin Cui Xun Suo |
| author_sort | Xinming Tang |
| collection | DOAJ |
| description | Abstract Eimeria tenella has emerged as valuable model organism for studying the biology and immunology of protozoan parasites with the establishment of the reverse genetic manipulation platform. In this report, we described the application of CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (endonuclease) system for efficient genetic editing in E. tenella, and showed that the CRISPR/Cas9 system mediates site-specific double-strand DNA breaks with a single guide RNA. Using this system, we successfully tagged the endogenous microneme protein 2 (EtMic2) by inserting the red fluorescent protein into the C-terminal of EtMic2. Our results extended the utility of the CRISPR/Cas9-mediated genetic modification system to E. tenella, and opened a new avenue for targeted investigation of gene functions in apicomplexan parasites. |
| first_indexed | 2024-12-19T22:10:39Z |
| format | Article |
| id | doaj.art-56f73fe75ce74f8c87a3e9a4ffea33eb |
| institution | Directory Open Access Journal |
| issn | 1297-9716 |
| language | English |
| last_indexed | 2024-12-19T22:10:39Z |
| publishDate | 2020-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Veterinary Research |
| spelling | doaj.art-56f73fe75ce74f8c87a3e9a4ffea33eb2022-12-21T20:03:55ZengBMCVeterinary Research1297-97162020-03-015111510.1186/s13567-020-00766-0Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 systemXinming Tang0Jingxia Suo1Lin Liang2Chunhui Duan3Dandan Hu4Xiaolong Gu5Yonglan Yu6Xianyong Liu7Shangjin Cui8Xun Suo9Institute of Animal Science, Chinese Academy of Agricultural SciencesKey Laboratory of Zoonosis of Ministry of Agriculture & National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural UniversityInstitute of Animal Science, Chinese Academy of Agricultural SciencesKey Laboratory of Zoonosis of Ministry of Agriculture & National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural UniversityKey Laboratory of Zoonosis of Ministry of Agriculture & National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural UniversityKey Laboratory of Zoonosis of Ministry of Agriculture & National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural UniversityKey Laboratory of Zoonosis of Ministry of Agriculture & National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural UniversityKey Laboratory of Zoonosis of Ministry of Agriculture & National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural UniversityInstitute of Animal Science, Chinese Academy of Agricultural SciencesKey Laboratory of Zoonosis of Ministry of Agriculture & National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural UniversityAbstract Eimeria tenella has emerged as valuable model organism for studying the biology and immunology of protozoan parasites with the establishment of the reverse genetic manipulation platform. In this report, we described the application of CRISPR (clustered regularly interspaced short palindromic repeat)/Cas9 (endonuclease) system for efficient genetic editing in E. tenella, and showed that the CRISPR/Cas9 system mediates site-specific double-strand DNA breaks with a single guide RNA. Using this system, we successfully tagged the endogenous microneme protein 2 (EtMic2) by inserting the red fluorescent protein into the C-terminal of EtMic2. Our results extended the utility of the CRISPR/Cas9-mediated genetic modification system to E. tenella, and opened a new avenue for targeted investigation of gene functions in apicomplexan parasites.http://link.springer.com/article/10.1186/s13567-020-00766-0 |
| spellingShingle | Xinming Tang Jingxia Suo Lin Liang Chunhui Duan Dandan Hu Xiaolong Gu Yonglan Yu Xianyong Liu Shangjin Cui Xun Suo Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system Veterinary Research |
| title | Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system |
| title_full | Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system |
| title_fullStr | Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system |
| title_full_unstemmed | Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system |
| title_short | Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system |
| title_sort | genetic modification of the protozoan eimeria tenella using the crispr cas9 system |
| url | http://link.springer.com/article/10.1186/s13567-020-00766-0 |
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