Korean Journal of Poultry Science (한국가금학회지)

The Korean Society of Poultry Science (한국가금학회)
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Non-Viral Transgenesis via Direct In Ovo Lipofection in Quail

비바이러스 In Ovo 직접주입법에 의한 메추리 형질전환 시스템

  • Park, Tae Sub (Graduate School of International Agricultural Technology and Institute of Green-Bio Science and Technology, Seoul National University) ;
  • Han, Jae Yong (Dept. of Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University)
  • 박태섭 (서울대학교 국제농업기술대학원) ;
  • 한재용 (서울대학교 농생명공학부)
  • Received : 2015.08.24
  • Accepted : 2015.09.07
  • Published : 2015.09.30
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Abstract

Transgenic animals have been widely used for developmental biology studies, as disease models, and even in industry such as transgenic bioreactor animals. For transgenic birds, quail has the great advantages of small body size, short generation time, and frequent egg production. To date, retroviral or lentiviral transduction has been used to generate transgenic quail for various purposes. However, the efficiency of transgenic offspring production with these methods is relatively low and viral vector usage has safety issues. Unfortunately, non-viral transgenesis has not been established in quail due to a deficiency of stem cell and germ cell culture systems. In this study, we established a direct in ovo lipofection method that could be used to create transgenic quail without germline-competent cells or viruses. To optimize the injection stage during embryo development, the liposome complex (containing piggyBacCMV-GFP and transposase plasmids) was introduced into an embryonic blood vessel at 50 hr, 55 hr or 60 hr. GFP expression was detected in various tissues (heart, kidney, liver and stomach) on day 12 of incubation under a fluorescence microscope. Additionally, GFP-positive cells were detected in the recipient embryonic gonads. In conclusion, the direct in ovo lipofection method with the piggyBac transposon could be an efficient and useful tool for generating transgenic quail.

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References

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