Generation of transgenic chicken through ovarian injection  

作　　者：Jinghua Jiang Caiying Wang Xuguang Du Fei Gao Sen Wu 

机构地区：[1]Frontiers Science Center for Molecular Design Breeding(MOE),State Key Laboratory of Animal Biotech Breeding,College of Biological Sciences,China Agricultural University,Beijing,China [2]Sanya Institute of China Agricultural University,Sanya,China

出　　处：《Animal Models and Experimental Medicine》2025年第1期187-193,共7页动物模型与实验医学(英文)

基　　金：National Key Research and Development Program of China,Grant/Award Number:2023YFF1000204;National Natural Science Foundation of China,Grant/Award Number:32172715,32230105 and 32330103;The Innovative Project of State Key Laboratory of Animal Biotech Breeding,Grant/Award Number:2024SKLAB1-2/9;Chinese Universities Scientific Fund,Grant/Award Number:2024TC167;The 2115 Talent Development Program of China Agricultural University。

摘　　要：Background : Traditional DNA microinjection methods used in mammals are difficult to apply to avian species due to their unique reproductive characteristics. Genetic manipulation in chickens, particularly involving immature follicles within living ovaries, has not been extensively explored. This study seeks to establish an efficient method for generating transgenic chickens through ovarian injection, potentially bypassing the challenges associated with primordial germ cell (PGC) manipulation and fertilized egg microinjection. Methods : Hens were anesthetized and underwent a surgical procedure to access the ovary for DNA injection into immature follicles. The study used liposomes to deliver GFP- expressing plasmids at various dosages. After injection, hens recovered, and their eggs were fertilized through artificial insemination. Results : Transgenic chickens were successfully generated in one generation without needing G0 founders. The injection of 20 μg plasmid yielded the highest transgenic efficiency at 12.1%. GFP- positive embryos were confirmed through microscopy, and successful transgene expression was validated at the tissue level using immunostaining. TERT and GFP elements introduced in the G1 generation resulted in 4.1% positive transgene rates, as confirmed by PCR and Southern blotting. Conclusion : This ovarian injection method offers a promising alternative for avian genetic manipulation, bypassing complex PGC procedures and enabling direct generation of G1 transgenic chickens. This technique simplifies the transgenic process for chickens and has the potential to be adapted for other avian species, especially those without established PGCs culture systems.

关 键 词：avian species gene manipulation ovarian injection transgenic chickens 

分 类 号：S83[农业科学—畜牧学]