Transgenic Chickens Expressing the 3D8 Single Chain Variable Fragment Protein Suppress Avian Influenza Transmission

Abstract The 3D8 single chain variable fragment (scFv) is a mini-antibody that causes unusual sequence-independent nuclease activity against all types of nucleic acids. We used recombinant lentiviruses to generate transgenic chickens expressing the 3D8 scFv gene under the control of the chicken β-ac...

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Autores principales:	Sung June Byun, Seong-su Yuk, Ye-Jin Jang, Hoonsung Choi, Mi-Hyang Jeon, TO Erdene-Ochir, Jung-Hoon Kwon, Jin-Yong Noh, Jeom Sun Kim, Jae Gyu YOO, Chang-Seon Song
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/73cef92afc1a48b7a251dd78228c0bb3
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spelling	oai:doaj.org-article:73cef92afc1a48b7a251dd78228c0bb32021-12-02T12:30:42ZTransgenic Chickens Expressing the 3D8 Single Chain Variable Fragment Protein Suppress Avian Influenza Transmission10.1038/s41598-017-05270-82045-2322https://doaj.org/article/73cef92afc1a48b7a251dd78228c0bb32017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05270-8https://doaj.org/toc/2045-2322Abstract The 3D8 single chain variable fragment (scFv) is a mini-antibody that causes unusual sequence-independent nuclease activity against all types of nucleic acids. We used recombinant lentiviruses to generate transgenic chickens expressing the 3D8 scFv gene under the control of the chicken β-actin promoter. From 420 injected embryos, 200 chicks (G0) hatched and were screened for the 3D8 scFv using PCR, and 15 chicks were identified as transgenic birds expressing the transgene in their semen. The G0 founder birds were mated with wild-type hens to produce seven transgenic chicks (G1). 3D8 scFv expression in the chicken embryonic fibroblasts (CEFs) was verified by RT-PCR and Western blot analysis. Immunofluorescence staining for 3D8 scFv in the CEFs revealed that the 3D8 scFv protein was primarily cytosolic. To identify 3D8 scFv anti-viral activity, wild-type and two transgenic CEF lines were infected with H9N2 avian influenza virus (AIV). We selected one line of transgenic chickens that exhibited the lowest number of plaque-forming units to be challenged with H9N2 virus. The challenge experiment revealed that contact exposed transgenic chickens expressing 3D8 scFv exhibited suppressed viral shedding. This results suggest that the transgenic chickens developed in this study could be useful for controlling potential within-flock AIV transmission.Sung June ByunSeong-su YukYe-Jin JangHoonsung ChoiMi-Hyang JeonTO Erdene-OchirJung-Hoon KwonJin-Yong NohJeom Sun KimJae Gyu YOOChang-Seon SongNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sung June Byun Seong-su Yuk Ye-Jin Jang Hoonsung Choi Mi-Hyang Jeon TO Erdene-Ochir Jung-Hoon Kwon Jin-Yong Noh Jeom Sun Kim Jae Gyu YOO Chang-Seon Song Transgenic Chickens Expressing the 3D8 Single Chain Variable Fragment Protein Suppress Avian Influenza Transmission
description	Abstract The 3D8 single chain variable fragment (scFv) is a mini-antibody that causes unusual sequence-independent nuclease activity against all types of nucleic acids. We used recombinant lentiviruses to generate transgenic chickens expressing the 3D8 scFv gene under the control of the chicken β-actin promoter. From 420 injected embryos, 200 chicks (G0) hatched and were screened for the 3D8 scFv using PCR, and 15 chicks were identified as transgenic birds expressing the transgene in their semen. The G0 founder birds were mated with wild-type hens to produce seven transgenic chicks (G1). 3D8 scFv expression in the chicken embryonic fibroblasts (CEFs) was verified by RT-PCR and Western blot analysis. Immunofluorescence staining for 3D8 scFv in the CEFs revealed that the 3D8 scFv protein was primarily cytosolic. To identify 3D8 scFv anti-viral activity, wild-type and two transgenic CEF lines were infected with H9N2 avian influenza virus (AIV). We selected one line of transgenic chickens that exhibited the lowest number of plaque-forming units to be challenged with H9N2 virus. The challenge experiment revealed that contact exposed transgenic chickens expressing 3D8 scFv exhibited suppressed viral shedding. This results suggest that the transgenic chickens developed in this study could be useful for controlling potential within-flock AIV transmission.
format	article
author	Sung June Byun Seong-su Yuk Ye-Jin Jang Hoonsung Choi Mi-Hyang Jeon TO Erdene-Ochir Jung-Hoon Kwon Jin-Yong Noh Jeom Sun Kim Jae Gyu YOO Chang-Seon Song
author_facet	Sung June Byun Seong-su Yuk Ye-Jin Jang Hoonsung Choi Mi-Hyang Jeon TO Erdene-Ochir Jung-Hoon Kwon Jin-Yong Noh Jeom Sun Kim Jae Gyu YOO Chang-Seon Song
author_sort	Sung June Byun
title	Transgenic Chickens Expressing the 3D8 Single Chain Variable Fragment Protein Suppress Avian Influenza Transmission
title_short	Transgenic Chickens Expressing the 3D8 Single Chain Variable Fragment Protein Suppress Avian Influenza Transmission
title_full	Transgenic Chickens Expressing the 3D8 Single Chain Variable Fragment Protein Suppress Avian Influenza Transmission
title_fullStr	Transgenic Chickens Expressing the 3D8 Single Chain Variable Fragment Protein Suppress Avian Influenza Transmission
title_full_unstemmed	Transgenic Chickens Expressing the 3D8 Single Chain Variable Fragment Protein Suppress Avian Influenza Transmission
title_sort	transgenic chickens expressing the 3d8 single chain variable fragment protein suppress avian influenza transmission
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/73cef92afc1a48b7a251dd78228c0bb3
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Transgenic Chickens Expressing the 3D8 Single Chain Variable Fragment Protein Suppress Avian Influenza Transmission

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