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

ABSTRACT 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 pr...

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Bibliographic Details
Published in:Poultry science 2019-01, Vol.98 (1), p.430-439
Main Authors: 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, Liu, Zhong-Hua, Mu, Yan-Shuang, Meng, Qing-Wen, Wang, Ning, Yang, Cai-Xia, Li, Hui
Format: Article
Language:English
Subjects:
Animals
Animals, Genetically Modified
Chickens - genetics
Embryo, Nonmammalian
Female
Gene Transfer Techniques - veterinary
Genetic Vectors
Green Fluorescent Proteins - genetics
Insemination, Artificial - veterinary
Male
Transfection - methods
Transfection - veterinary
Transgenes
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Summary:ABSTRACT 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 G0 transgenic roosters. From 27 G0 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 G0 rooster with the highest number of EGFP fluorescent sperm, one G1 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.
ISSN:0032-5791
1525-3171
DOI:10.3382/ps/pey321