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

Collection with item attached
2017
Item details URL
http://open-repository.kisti.re.kr/cube/handle/open_repository/486443.do
DOI
10.1038/s41598-017-05270-8
Title
Transgenic Chickens Expressing the 3D8 Single Chain Variable FragmentProtein Suppress Avian Influenza Transmission
Description
We are grateful to Ha-Na Youn, Hyo-Seon Joo, Kyung-Min Kim, and HeenariKoo for their excellent technical support. This work was conducted withthe support of the "Cooperative Research Program for Agriculture Science& Technology Development (Project No. PJ00646904)" Rural DevelopmentAdministration, Republic of Korea. This research was partly supported bythe Konkuk University Research Program (2014).
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 beta-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.
provenance
Made available in Cube on 2018-09-28T16:07:46Z (GMT). No. of bitstreams: 0
language
English
author
Byun, Sung June
Yuk, Seong-su
Jang, Ye-Jin
Choi, Hoonsung
Jeon, Mi-Hyang
Erdene-Ochir, To
Kwon, Jung-Hoon
Noh, Jin-Yong
Kim, Jeom Sun
Yoo, Jae Gyu
Song, Chang-Seon
orcid
Yoo, Jae Gyu/0000-0002-8542-9193
accessioned
2018-09-28T16:07:46Z
available
2018-09-28T16:07:46Z
issued
2017
citation
SCIENTIFIC REPORTS(7)
issn
2045-2322
uri
http://open-repository.kisti.re.kr/cube/handle/open_repository/486443.do
Funder
농촌진흥청
Funding Program
축산시험연구(R&D,책임운영)
Project ID
1395030951
Jurisdiction
Rep.of Korea
Project Name
Development of technique for avian influenza resistant chicken production
rights
openAccess
type
article


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