Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.

Although different strategies have been developed to generate transgenic poultry, low efficiency of germline transgene transmission has remained a challenge in poultry transgenesis. Herein, we developed an efficient germline transgenesis method using a lentiviral vector system in chickens through mu...

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Autores principales:	Danial Naseri, Kianoush Dormiani, Mehdi Hajian, Farnoosh Jafarpour, Mahboobeh Forouzanfar, Naeimeh Karimi, Mohammad Hossein Nasr-Esfahani
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/be0029e3b9f54f6c84c2d4fb67354f8b
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spelling	oai:doaj.org-article:be0029e3b9f54f6c84c2d4fb67354f8b2021-12-02T20:03:56ZImproving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.1932-620310.1371/journal.pone.0247471https://doaj.org/article/be0029e3b9f54f6c84c2d4fb67354f8b2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0247471https://doaj.org/toc/1932-6203Although different strategies have been developed to generate transgenic poultry, low efficiency of germline transgene transmission has remained a challenge in poultry transgenesis. Herein, we developed an efficient germline transgenesis method using a lentiviral vector system in chickens through multiple injections of transgenes into embryos at different stages of development. The embryo chorioallantoic membrane (CAM) vasculature was successfully used as a novel route of gene transfer into germline tissues. Compared to the other routes of viral vector administration, the embryo's bloodstream at Hamburger-Hamilton (HH) stages 14-15 achieved the highest rate of germline transmission (GT), 7.7%. Single injection of viral vectors into the CAM vasculature resulted in a GT efficiency of 2.7%, which was significantly higher than the 0.4% obtained by injection into embryos at the blastoderm stage. Double injection of viral vectors into the bloodstream at HH stages 14-15 and through CAM was the most efficient method for producing germline chimeras, giving a GT rate of 13.6%. The authors suggest that the new method described in this study could be efficiently used to produce transgenic poultry in virus-mediated gene transfer systems.Danial NaseriKianoush DormianiMehdi HajianFarnoosh JafarpourMahboobeh ForouzanfarNaeimeh KarimiMohammad Hossein Nasr-EsfahaniPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 6, p e0247471 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Danial Naseri Kianoush Dormiani Mehdi Hajian Farnoosh Jafarpour Mahboobeh Forouzanfar Naeimeh Karimi Mohammad Hossein Nasr-Esfahani Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.
description	Although different strategies have been developed to generate transgenic poultry, low efficiency of germline transgene transmission has remained a challenge in poultry transgenesis. Herein, we developed an efficient germline transgenesis method using a lentiviral vector system in chickens through multiple injections of transgenes into embryos at different stages of development. The embryo chorioallantoic membrane (CAM) vasculature was successfully used as a novel route of gene transfer into germline tissues. Compared to the other routes of viral vector administration, the embryo's bloodstream at Hamburger-Hamilton (HH) stages 14-15 achieved the highest rate of germline transmission (GT), 7.7%. Single injection of viral vectors into the CAM vasculature resulted in a GT efficiency of 2.7%, which was significantly higher than the 0.4% obtained by injection into embryos at the blastoderm stage. Double injection of viral vectors into the bloodstream at HH stages 14-15 and through CAM was the most efficient method for producing germline chimeras, giving a GT rate of 13.6%. The authors suggest that the new method described in this study could be efficiently used to produce transgenic poultry in virus-mediated gene transfer systems.
format	article
author	Danial Naseri Kianoush Dormiani Mehdi Hajian Farnoosh Jafarpour Mahboobeh Forouzanfar Naeimeh Karimi Mohammad Hossein Nasr-Esfahani
author_facet	Danial Naseri Kianoush Dormiani Mehdi Hajian Farnoosh Jafarpour Mahboobeh Forouzanfar Naeimeh Karimi Mohammad Hossein Nasr-Esfahani
author_sort	Danial Naseri
title	Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.
title_short	Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.
title_full	Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.
title_fullStr	Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.
title_full_unstemmed	Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.
title_sort	improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/be0029e3b9f54f6c84c2d4fb67354f8b
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Improving germline transmission efficiency in chimeric chickens using a multi-stage injection approach.

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