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Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system

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)...

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Detalles Bibliográficos
Autores principales: Tang, Xinming, Suo, Jingxia, Liang, Lin, Duan, Chunhui, Hu, Dandan, Gu, Xiaolong, Yu, Yonglan, Liu, Xianyong, Cui, Shangjin, Suo, Xun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7065449/
https://www.ncbi.nlm.nih.gov/pubmed/32160917
http://dx.doi.org/10.1186/s13567-020-00766-0
Descripción
Sumario: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.