Exploring transgene transfer from the transgenic chicken model to its offspring through a nonviral vector

Pan Lin Wang; Xue Qin Lei; Ting Sheng Xu; Ming Yan Shi; Zhen Song; Zhen Hong Li; Guang Hui Wei; Guang Ping Zhang; Jun Tang Li

doi:10.1134/S1068162017050156

Exploring transgene transfer from the transgenic chicken model to its offspring through a nonviral vector

Authors: Wang P.L.¹, Lei X.Q.¹, Xu T.S.¹, Shi M.Y.², Song Z.¹, Li Z.H.¹, Wei G.H.³, Zhang G.P.⁴, Li J.T.⁵
Affiliations:
1. College of Animal Science and Technology
2. College of Life Science
3. College of Animal Science and Veterinary Medicine
4. Henan University of Science and Technology Affiliated First Hospital
5. Centre of Inflammation and Cancer Research
Issue: Vol 43, No 5 (2017)
Pages: 570-575
Section: Article
URL: https://journals.rcsi.science/1068-1620/article/view/228674
DOI: https://doi.org/10.1134/S1068162017050156
ID: 228674

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Abstract

There is much interest in using chickens as “bioreactors” to produce large quantities of biopharmaceuticals. However, transient expression of foreign genes have been known to cause low efficiency of obtaining transgenic offspring, especially when using nonviral vectors. In present study, a transgenic chicken model was investigated to determine whether an exogenous gene can be expressed stably and transferred to its offspring through a matrix attachment region (MAR)-mediated non-viral vector using the eGFP marker gene. The eukaryotic expression vector pEGFP-N1-MAR, which contains the eGFP gene and MAR, was constructed and transfected into a chicken stage-X blastoderm to produce a G0 generation of transgenic chickens. The hatchabilities of different injection regions were tested; 18 of the 40 eggs injected with pEGFPN1- MAR in the area opaca hatched after 21 days of incubation, and had a hatchability rate of 45%. By contrast, eggs injected at the area pellucida did not hatch. Results from the fluorescence signal detection and polymerase chain reaction (PCR) verified that four hatched chicks from the G0 generation expressed the eGFP gene. Furthermore, fluorescence signal detection results indicated that 2 of the 65 chicks from the G1 generation expressed the eGFP gene. We conclude that MAR facilitates the production of transgenic chickens; pEGFP-N1-MAR application is a novel approach that can produce transgenic chicken offspring.

Keywords

transgenic chicken, bioreactor, matrix attachment region, nonviral vector, area opaca, area pellucida

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Exploring transgene transfer from the transgenic chicken model to its offspring through a nonviral vector

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