Production of transgenic broilers by non-viral vectors via optimizing egg windowing and screening transgenic roosters

文献类型： 外文期刊

第一作者： Wang, Zhong-Bin

作者： Wang, Zhong-Bin;Du, Zhi-Qiang;Na, Wei;Jing, Jun-Hong;Li, Yu-Mao;Leng, Li;Luan, Peng;Wu, Chun-Yan;Zhang, Ke;Wang, Yu-Xiang;Liu, Wen-Li;Yuan, Hui;Wang, Ning;Yang, Cai-Xia;Li, Hui;Wang, Zhong-Bin;Du, Zhi-Qiang;Na, Wei;Jing, Jun-Hong;Li, Yu-Mao;Leng, Li;Luan, Peng;Wu, Chun-Yan;Zhang, Ke;Wang, Yu-Xiang;Liu, Wen-Li;Yuan, Hui;Wang, Ning;Yang, Cai-Xia;Li, Hui;Wang, Zhong-Bin;Du, Zhi-Qiang;Na, Wei;Jing, Jun-Hong;Li, Yu-Mao;Leng, Li;Luan, Peng;Wu, Chun-Yan;Zhang, Ke;Wang, Yu-Xiang;Liu, Wen-Li;Yuan, Hui;Wang, Ning;Yang, Cai-Xia;Li, Hui;Liu, Zhong-Hua;Mu, Yan-Shuang;Meng, Qing-Wen

作者机构：

关键词： egg windowing; in ovo transfection; EGFP sperm; transgenic chicken

期刊名称：POULTRY SCIENCE （ 影响因子：3.352； 五年影响因子：3.679 ）

ISSN： 0032-5791

年卷期： 2019 年 98 卷 1 期

页码：

收录情况： SCI

摘要： The generation of transgenic chickens is of both biomedical and agricultural significance, and recently chicken transgenesis technology has been greatly advanced. However, major issues still exist in the efficient production of transgenic chickens. This study was designed to optimize the production of enhanced green fluorescence protein (EGFP)-transgenic broilers, including egg windowing at the blunt end (air cell) of egg, and the direct transfection of circulating primordial germ cells by microinjection of the Tol2 plasmid-liposome complex into the early embryonic dorsal aorta. For egg windowing, we discovered that proper manipulation of the inner shell membrane at the blunt end could improve the rate of producing G(0) transgenic roosters. From 27 G(0) roosters, we successfully collected semen with EGFP-positive sperms from 16 and 19 roosters after direct fluorescence observation and fluorescence-activated cell sorting analyses (13 detected by both methods), respectively. After artificial insemination using the G(0) rooster with the highest number of EGFP fluorescent sperm, one G(1) EGFP transgenic broiler (1/81, 1.23%) was generated. Our results indicate that appropriate egg windowing and screening of potentially transgene-positive roosters can improve the production of germline-transmitted transgenic birds.

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