Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system
Recent advances in the study of the CRISPR/Cas9 system have provided a precise and versatile approach for genome editing in various species. However, the applicability and efficiency of this method in large animal models, such as the goat, have not been extensively studied. Here, by co-injection of...
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| Format: | Online |
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Nature Publishing Group
2015
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4564737/ |
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pubmed-45647372015-09-15 Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system Wang, Xiaolong Yu, Honghao Lei, Anmin Zhou, Jiankui Zeng, Wenxian Zhu, Haijing Dong, Zhiming Niu, Yiyuan Shi, Bingbo Cai, Bei Liu, Jinwang Huang, Shuai Yan, Hailong Zhao, Xiaoe Zhou, Guangxian He, Xiaoling Chen, Xiaoxu Yang, Yuxin Jiang, Yu Shi, Lei Tian, Xiue Wang, Yongjun Ma, Baohua Huang, Xingxu Qu, Lei Chen, Yulin Article Recent advances in the study of the CRISPR/Cas9 system have provided a precise and versatile approach for genome editing in various species. However, the applicability and efficiency of this method in large animal models, such as the goat, have not been extensively studied. Here, by co-injection of one-cell stage embryos with Cas9 mRNA and sgRNAs targeting two functional genes (MSTN and FGF5), we successfully produced gene-modified goats with either one or both genes disrupted. The targeting efficiency of MSTN and FGF5 in cultured primary fibroblasts was as high as 60%, while the efficiency of disrupting MSTN and FGF5 in 98 tested animals was 15% and 21% respectively, and 10% for double gene modifications. The on- and off-target mutations of the target genes in fibroblasts, as well as in somatic tissues and testis of founder and dead animals, were carefully analyzed. The results showed that simultaneous editing of several sites was achieved in large animals, demonstrating that the CRISPR/Cas9 system has the potential to become a robust and efficient gene engineering tool in farm animals, and therefore will be critically important and applicable for breeding. Nature Publishing Group 2015-09-10 /pmc/articles/PMC4564737/ /pubmed/26354037 http://dx.doi.org/10.1038/srep13878 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Wang, Xiaolong Yu, Honghao Lei, Anmin Zhou, Jiankui Zeng, Wenxian Zhu, Haijing Dong, Zhiming Niu, Yiyuan Shi, Bingbo Cai, Bei Liu, Jinwang Huang, Shuai Yan, Hailong Zhao, Xiaoe Zhou, Guangxian He, Xiaoling Chen, Xiaoxu Yang, Yuxin Jiang, Yu Shi, Lei Tian, Xiue Wang, Yongjun Ma, Baohua Huang, Xingxu Qu, Lei Chen, Yulin |
| spellingShingle |
Wang, Xiaolong Yu, Honghao Lei, Anmin Zhou, Jiankui Zeng, Wenxian Zhu, Haijing Dong, Zhiming Niu, Yiyuan Shi, Bingbo Cai, Bei Liu, Jinwang Huang, Shuai Yan, Hailong Zhao, Xiaoe Zhou, Guangxian He, Xiaoling Chen, Xiaoxu Yang, Yuxin Jiang, Yu Shi, Lei Tian, Xiue Wang, Yongjun Ma, Baohua Huang, Xingxu Qu, Lei Chen, Yulin Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system |
| author_facet |
Wang, Xiaolong Yu, Honghao Lei, Anmin Zhou, Jiankui Zeng, Wenxian Zhu, Haijing Dong, Zhiming Niu, Yiyuan Shi, Bingbo Cai, Bei Liu, Jinwang Huang, Shuai Yan, Hailong Zhao, Xiaoe Zhou, Guangxian He, Xiaoling Chen, Xiaoxu Yang, Yuxin Jiang, Yu Shi, Lei Tian, Xiue Wang, Yongjun Ma, Baohua Huang, Xingxu Qu, Lei Chen, Yulin |
| author_sort |
Wang, Xiaolong |
| title |
Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system |
| title_short |
Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system |
| title_full |
Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system |
| title_fullStr |
Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system |
| title_full_unstemmed |
Generation of gene-modified goats targeting MSTN and FGF5 via zygote injection of CRISPR/Cas9 system |
| title_sort |
generation of gene-modified goats targeting mstn and fgf5 via zygote injection of crispr/cas9 system |
| description |
Recent advances in the study of the CRISPR/Cas9 system have provided a precise and versatile approach for genome editing in various species. However, the applicability and efficiency of this method in large animal models, such as the goat, have not been extensively studied. Here, by co-injection of one-cell stage embryos with Cas9 mRNA and sgRNAs targeting two functional genes (MSTN and FGF5), we successfully produced gene-modified goats with either one or both genes disrupted. The targeting efficiency of MSTN and FGF5 in cultured primary fibroblasts was as high as 60%, while the efficiency of disrupting MSTN and FGF5 in 98 tested animals was 15% and 21% respectively, and 10% for double gene modifications. The on- and off-target mutations of the target genes in fibroblasts, as well as in somatic tissues and testis of founder and dead animals, were carefully analyzed. The results showed that simultaneous editing of several sites was achieved in large animals, demonstrating that the CRISPR/Cas9 system has the potential to become a robust and efficient gene engineering tool in farm animals, and therefore will be critically important and applicable for breeding. |
| publisher |
Nature Publishing Group |
| publishDate |
2015 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4564737/ |
| _version_ |
1613474287703818240 |