Transgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.

Several strategies have been used to generate transgenic birds. The most successful method so far has been the injection of lentiviral vectors into the subgerminal cavity of a newly laid egg. We report here a new, easy and effective way to produce transgenic quails through direct injection of a lent...

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Autores principales:	Zifu Zhang, Peng Sun, Fuxian Yu, Li Yan, Fang Yuan, Wenxin Zhang, Tao Wang, Zhiyi Wan, Qiang Shao, Zandong Li
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/21669a1dec8748a6a7f6b79e622d8ad3
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spelling	oai:doaj.org-article:21669a1dec8748a6a7f6b79e622d8ad32021-11-18T08:05:27ZTransgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.1932-620310.1371/journal.pone.0050817https://doaj.org/article/21669a1dec8748a6a7f6b79e622d8ad32012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23251391/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Several strategies have been used to generate transgenic birds. The most successful method so far has been the injection of lentiviral vectors into the subgerminal cavity of a newly laid egg. We report here a new, easy and effective way to produce transgenic quails through direct injection of a lentiviral vector, containing an enhanced-green fluorescent protein (eGFP) transgene, into the blood vessels of quail embryos at Hamburger-Hamilton stage 13-15 (HH13-15). A total of 80 embryos were injected and 48 G0 chimeras (60%) were hatched. Most injected embryo organs and tissues of hatched quails were positive for eGFP. In five out of 21 mature G0 male quails, the semen was eGFP-positive, as detected by polymerase chain reaction (PCR), indicating transgenic germ line chimeras. Testcross and genetic analyses revealed that the G0 quail produced transgenic G1 offspring; of 46 G1 hatchlings, 6 were transgenic (6/46, 13.0%). We also compared this new method with the conventional transgenesis using stage X subgerminal cavity injection. Total 240 quail embryos were injected by subgerminal cavity injection, of which 34 (14.1%) were hatched, significantly lower than the new method. From these hatched quails semen samples were collected from 19 sexually matured males and tested for the transgene by PCR. The transgene was present in three G0 male quails and only 4/236 G1 offspring (1.7%) were transgenic. In conclusion, we developed a novel bird transgenic method by injection of lentiviral vector into embryonic blood vessel at HH 13-15 stage, which result in significant higher transgenic efficiency than the conventional subgerminal cavity injection.Zifu ZhangPeng SunFuxian YuLi YanFang YuanWenxin ZhangTao WangZhiyi WanQiang ShaoZandong LiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e50817 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Zifu Zhang Peng Sun Fuxian Yu Li Yan Fang Yuan Wenxin Zhang Tao Wang Zhiyi Wan Qiang Shao Zandong Li Transgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.
description	Several strategies have been used to generate transgenic birds. The most successful method so far has been the injection of lentiviral vectors into the subgerminal cavity of a newly laid egg. We report here a new, easy and effective way to produce transgenic quails through direct injection of a lentiviral vector, containing an enhanced-green fluorescent protein (eGFP) transgene, into the blood vessels of quail embryos at Hamburger-Hamilton stage 13-15 (HH13-15). A total of 80 embryos were injected and 48 G0 chimeras (60%) were hatched. Most injected embryo organs and tissues of hatched quails were positive for eGFP. In five out of 21 mature G0 male quails, the semen was eGFP-positive, as detected by polymerase chain reaction (PCR), indicating transgenic germ line chimeras. Testcross and genetic analyses revealed that the G0 quail produced transgenic G1 offspring; of 46 G1 hatchlings, 6 were transgenic (6/46, 13.0%). We also compared this new method with the conventional transgenesis using stage X subgerminal cavity injection. Total 240 quail embryos were injected by subgerminal cavity injection, of which 34 (14.1%) were hatched, significantly lower than the new method. From these hatched quails semen samples were collected from 19 sexually matured males and tested for the transgene by PCR. The transgene was present in three G0 male quails and only 4/236 G1 offspring (1.7%) were transgenic. In conclusion, we developed a novel bird transgenic method by injection of lentiviral vector into embryonic blood vessel at HH 13-15 stage, which result in significant higher transgenic efficiency than the conventional subgerminal cavity injection.
format	article
author	Zifu Zhang Peng Sun Fuxian Yu Li Yan Fang Yuan Wenxin Zhang Tao Wang Zhiyi Wan Qiang Shao Zandong Li
author_facet	Zifu Zhang Peng Sun Fuxian Yu Li Yan Fang Yuan Wenxin Zhang Tao Wang Zhiyi Wan Qiang Shao Zandong Li
author_sort	Zifu Zhang
title	Transgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.
title_short	Transgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.
title_full	Transgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.
title_fullStr	Transgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.
title_full_unstemmed	Transgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.
title_sort	transgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/21669a1dec8748a6a7f6b79e622d8ad3
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Transgenic quail production by microinjection of lentiviral vector into the early embryo blood vessels.

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