Disruption of the sheep BMPR-IB gene by CRISPR/Cas9 in in vitro-produced embryos

文献类型： 外文期刊

第一作者： Zhang, Xuemei

作者： Zhang, Xuemei;Li, Wenrong;Liu, Mingjun;Zhang, Xuemei;Li, Wenrong;Wu, Yangsheng;Peng, Xinrong;Lou, Bian;Wang, Liqin;Liu, Mingjun;Zhang, Xuemei;Li, Wenrong;Wu, Yangsheng;Peng, Xinrong;Lou, Bian;Wang, Liqin;Liu, Mingjun

作者机构：

关键词： Sheep;BMPR-IB;CRISPR/Cas9;Embryo;Gene editing;Indel

期刊名称：THERIOGENOLOGY （ 影响因子：2.74； 五年影响因子：2.93 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： BMPR-IB (also known as FecB) is a key candidate gene for the genetic control of sheep reproductive performance. Loss-of-function mutations in the sheep BMPR-IB gene lead to an increase in ovulation rate and consequently larger litter size. However, the BMPR-IB gene has been identified in only a few sheep breeds. To improve sheep reproduction through modification of the BMPR-IB gene, we designed an sgRNA to target the sheep BMPR-IB gene by using the CRISPR/Cas9 system. First, we performed gene editing by injecting Cas9/sgRNA into the cytoplasm of one-cell fertilized eggs. A total of 88 embryos were assayed by T7EI digestion and Sanger sequencing. The results reported that the efficiency of gene modification was 37.5% (33/88) and increased with the developmental stage of embryo from the 2-cell stage to the blastocyst stage. Of the 33 gene editing embryos, 12 (36%, 12/33) were homozygous and 21 (64%, 21/33) were heterozygous. Moreover, sequence analysis of the PCR products from the positive embryos revealed that there were more than 10 modification forms that resulted in frame shift and truncated proteins. Further analysis by cloning and sequencing of each individual embryo showed a high level of mosaicism. In addition, off-target event analysis revealed that none of the off target mutations was introduced into the embryos. Our results indicated that the Cas9/sgRNA system is a simple and efficient tool that may potentially be used in the genetic modification of the sheep BMPR-IB gene in vivo. (C) 2016 Elsevier Inc. All rights reserved.

分类号： Q95

  • 相关文献

[1]Long-term cryopreservation had no adverse effect on viability of embryos and their offspring in sheep. Yao, Yu-Chang,Li, Wu,Qi, Mei-Yu,Lu, Ming -Hai,Wang, Su-Mei,Han, Hong-Bing. 2012

[2]CRISPR/Cas9-mediated ApoE(-/-) and LDLR-/- double gene knockout in pigs elevates serum LDL-C and TC levels. Huang, Lei,Cheng, Ying,Xu, Kui,Gao, Qian,Xia, Ying,Liu, Yang,Zhang, Xue,Mu, Yulian,Li, Kui,Hua, Zaidong,Xiao, Hongwei,Zheng, Xinming,Huang, Lei. 2017

[3]A Study on BMPR-IB Genes of Bayanbulak Sheep. Zuo, Beiyao,Wang, Ziyu,Ying, Shijia,Wang, Feng,Zuo, Beiyao,Wang, Xu,Nisa, Noor,Hu, Xiaolong,Gong, Changhai,Qian, Hongguan,Bayier, Aierdin,Guo, Zhiqin.

[4]Identification of Sesame Genomic Variations from Genome Comparison of Landrace and Variety. Wei, Xin,Zhu, Xiaodong,Yu, Jingyin,Wang, Linhai,Zhang, Yanxin,Li, Donghua,Zhou, Rong,Zhang, Xiurong. 2016

[5]Development of novel InDel markers and genetic diversity in Chenopodium quinoa through whole-genome re-sequencing. Zhang, Tifu,Lv, Yuanda,Zhou, Ling,Lu, Haiyan,Liang, Shuaiqiang,Bao, Huabin,Zhao, Han,Gu, Minfeng,Liu, Yuhe. 2017

[6]Association Mapping Reveals Genetic Loci Associated with Important Agronomic Traits in Lentinula edodes, Shiitake Mushroom. Li, Chuang,Gong, Wenbing,Zhang, Lin,Bian, Yinbing,Xiao, Yang,Gong, Wenbing,Yang, Zhiquan,Nong, Wenyan,Kwan, Hoi-Shan,Cheung, Man-Kit. 2017

[7]Development of a large number of SSR and InDel markers and construction of a high-density genetic map based on a RIL population of pepper (Capsicum annuum L.). Zhang, Xiao-fen,Wang, Qian,Zhang, Xiao-fen,Sun, Hong-he,Xu, Yong,Chen, Bin,Yu, Shuan-cang,Geng, San-sheng.

[8]Development of InDel markers for the restorer gene and Rf1 assessment of their utility for marker-assisted selection in cotton. Jianyong Wu,Xing, Chaozhu,Meng Zhang,Xuexian Zhang,Liping Guo,Tingxiang Qi,Hailin Wang,Huini Tang,Jinfa Zhang,Chaozhu Xing.

[9]Genetic analysis and molecular characterization of Chinese sesame (Sesamum indicum L.) cultivars using Insertion-Deletion (InDel) and Simple Sequence Repeat (SSR) markers. Wu, Kun,Yang, Minmin,Liu, Hongyan,Mei, Ju,Zhao, Yingzhong,Tao, Ye. 2014

[10]Development of 1047 insertion-deletion markers for rice genetic studies and breeding. Zeng, Y. X.,Wen, Z. H.,Ma, L. Y.,Ji, Z. J.,Li, X. M.,Yang, C. D.. 2013

[11]Genome re-sequencing of two accessions and fine mapping the locus of lobed leaflet margins in mungbean. Jiao, Keyuan,Guo, Wuxiu,Jiao, Keyuan,Li, Xin,Yuan, Xingxing,Yuan, Xingxing,Cui, Xiaoyan,Chen, Xin.

[12]Exploiting Illumina sequencing for the development of InDel markers in watermelon (Citrullus lanatus). Liu, G.,Xu, J. H.,Zhang, M.,Li, P. F.,Yao, X. F.,Hou, Q.,Zhu, L. L.,Ren, R. S.,Yang, X. P..

[13]Creation of targeted inversion mutations in plants using an RNA-guided endonuclease. Zhang, Congsheng,Liu, Changlin,Weng, Jianfeng,Liu, Fang,Li, Xinhai,Xie, Chuanxiao,Zhang, Congsheng,Cheng, Beijiu. 2017

[14]Knock out of the annexin gene OsAnn3 via CRISPR/Cas9-mediated genome editing decreased cold tolerance in rice. Tang, Ning,Cao, Mengliang,Shen, Chunxiu,Que, Zhiqun,Cao, Mengliang,Li, Ding,He, Ronghua. 2017

[15]Gene targeting using the Agrobacterium tumefaciens-mediated CRISPR-Cas system in rice. Xu, Rongfang,Li, Hao,Qin, Ruiying,Wang, Lu,Li, Li,Wei, Pengcheng,Yang, Jianbo,Li, Hao,Wei, Pengcheng,Xu, Rongfang,Li, Li. 2014

[16]A Simple and Efficient Method for CRISPR/Cas9-Induced Rice Mutant Screening. Feng Xu-ping,Zhang Chu,Liu Xiao-dan,Shen Ting-ting,He Yong,Feng Xu-ping,Zhang Chu,Liu Xiao-dan,Shen Ting-ting,He Yong,Peng Cheng,Xu Jun-feng. 2018

[17]Deletion of the Bombyx mori odorant receptor co-receptor (BmOrco) impairs olfactory sensitivity in silkworms. Hao, Dejun,Liu, Qun,Hao, Dejun,Liu, Qun,Zeng, Baosheng,Huang, Yongping,Wang, Guirong.

[18]Production of alpha 1,3-galactosyltransferase and cytidine monophosphate-N-acetylneuraminic acid hydroxylase gene double-deficient pigs by CRISPR/Cas9 and handmade cloning. Gao, Hanchao,Zhao, Chengjiang,Zhao, Yanli,Li, Zesong,Cai, Zhiming,Mou, Lisha,Gao, Hanchao,Zhao, Chengjiang,Xiang, Xi,Li, Yong,Pan, Dengke,Dai, Yifan,Hara, Hidetaka,Cooper, David K. C..

[19]Targeted mutagenesis by CRISPR/Cas9 system in the model legume Medicago truncatula. Meng, Yingying,Hou, Yaling,Wang, Hui,Niu, Lifang,Lin, Hao,Ji, Ronghuan,Liu, Bin,Wen, Jiangqi.

[20]Live attenuated pseudorabies virus developed using the CRISPR/Cas9 system. Tang, Yan-Dong,Liu, Ji-Ting,Wang, Tong-Yun,An, Tong-Qing,Sun, Ming-Xia,Wang, Shu-Jie,Fang, Qiong-Qiong,Hou, Lin-lin,Tian, Zhi-Jun,Cai, Xue-Hui,Liu, Ji-Ting.

作者其他论文 更多>>

微信小程序