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RAPID COMMUNICATION : Generation of FGF5 knockout sheep via the CRISPR/Cas9 system

文献类型： 外文期刊

作者： Hu, R. 1 ; Fan, Z. Y. 1 ; Wang, B. Y. 2 ; Deng, S. L. 3 ; Zhang, X. S. 4 ; Zhang, J. L. 4 ; Han, H. B. 1 ; Lian, Z. X. 1 ;

作者机构： 1.China Agr Univ, Beijing Key Lab Anim Genet Improvement, Beijing, Peoples R China

2.Chinese Acad Agr Sci, Inst Anim Sci, Beijing, Peoples R China

3.Chinese Acad Sci, Inst Zool, State Key Lab Reprod Biol, Beijing 100101, Peoples R China

4.Tianjin Acad Agr Sci, Inst Anim Sci, Tianjin, Peoples R China

关键词： Cas9;fibroblast growth factor 5;knockout;sheep;wool

期刊名称：JOURNAL OF ANIMAL SCIENCE （ 影响因子：3.159； 五年影响因子：2.912 ）

ISSN： 0021-8812

年卷期： 2017 年 95 卷 5 期

页码：

收录情况： SCI

摘要： Sheep are an important source of fiber production. Fibroblast growth factor 5 (FGF5) is a dominant inhibitor of length of the anagen phase of the hair cycle. Knockout or silencing of the FGF5 gene results in a wooly coat in mice, donkeys, dogs, and rabbits. In sheep breeding, wool length is one of the most important wool quality traits. However, traditional breeding cannot accurately and efficiently mediate an advanced genotype into the sheep genome. In this study, we generated 3 FGF5 knockout sheep via the 1-step clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system. Sequencing analysis confirmed that mutations in the FGF5 gene existed in all germ lines of 3 founders: besides the intact sequence, 3 kinds of deletions in the FGF5 gene (including 5, 13, and 33 bp) were detected. The changes in the primary and senior structure of the FGF5 protein due to the 3 deletions in founders suggested that the FGF5 protein was dysfunctional. In addition, the expression level of intact FGF5 mRNA in heterozygous individuals decreased compared with the wild types (P < 0.01). Functionally, we discovered that wool length in founders was significantly longer than in wild types (P < 0.05). Collectively, the FGF5 knockout sheep with the longer wool length phenotype will provide an efficient way for fast genetic improvement of sheep breeding and promote the development of wool industry.

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