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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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:
Short Report
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
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author Tang, Xinming
Suo, Jingxia
Liang, Lin
Duan, Chunhui
Hu, Dandan
Gu, Xiaolong
Yu, Yonglan
Liu, Xianyong
Cui, Shangjin
Suo, Xun
author_facet Tang, Xinming
Suo, Jingxia
Liang, Lin
Duan, Chunhui
Hu, Dandan
Gu, Xiaolong
Yu, Yonglan
Liu, Xianyong
Cui, Shangjin
Suo, Xun
author_sort Tang, Xinming
collection PubMed
description 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.
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spelling pubmed-70654492020-03-16 Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system Tang, Xinming Suo, Jingxia Liang, Lin Duan, Chunhui Hu, Dandan Gu, Xiaolong Yu, Yonglan Liu, Xianyong Cui, Shangjin Suo, Xun Vet Res Short Report 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. BioMed Central 2020-03-11 2020 /pmc/articles/PMC7065449/ /pubmed/32160917 http://dx.doi.org/10.1186/s13567-020-00766-0 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Short Report
Tang, Xinming
Suo, Jingxia
Liang, Lin
Duan, Chunhui
Hu, Dandan
Gu, Xiaolong
Yu, Yonglan
Liu, Xianyong
Cui, Shangjin
Suo, Xun
Genetic modification of the protozoan Eimeria tenella using the CRISPR/Cas9 system
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
topic Short Report
url 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
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