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Generation of a transgenic pig expressing human dipeptidylpeptidase-4 (DPP-4)

Human dipeptidylpeptidase-4(DPP-4) 발현 형질전환 돼지의 생산

  • Chung, Hak Jae (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Sa, Soo Jin (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Baek, Sun Young (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Cho, Eun Suek (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Young Shin (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Hong, Jun Ki (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Cho, Kyu Ho (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Ji Youn (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Park, Mi Ryung (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Kyung Woon (Planning & Coordination Division, National Institute of Animal Science, Rural Development Administration)
  • 정학재 (농촌진흥청 국립축산과학원 양돈과) ;
  • 사수진 (농촌진흥청 국립축산과학원 양돈과) ;
  • 백선영 (농촌진흥청 국립축산과학원 양돈과) ;
  • 조은석 (농촌진흥청 국립축산과학원 양돈과) ;
  • 김영신 (농촌진흥청 국립축산과학원 양돈과) ;
  • 홍준기 (농촌진흥청 국립축산과학원 양돈과) ;
  • 조규호 (농촌진흥청 국립축산과학원 양돈과) ;
  • 김지윤 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 박미령 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 김경운 (농촌진흥청 국립축산과학원 기획조정과)
  • Received : 2019.07.02
  • Accepted : 2019.09.06
  • Published : 2019.09.30
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Abstract

As dipeptidyl peptidase-4(DPP-4) inhibitors are used widely as a secondary treatment for type 2 diabetes because they tend to be well tolerated with minimal side effects, the human DPP-4(hDPP-4) gene was injected into a pig zygote through micro-injection, and 1-cell stage fertilized embryos were then transplanted surgically into the oviduct. Three pigs were fertilized with hDPP-4 genes and produced sixteen piglets, in which one male piglet was identified to be transgenic. Finally, transgenic pigs showing hDPP-4 gene expression in the tail were produced. Western blot and RT-PCR analysis confirmed that the hDPP-4 is expressed strongly in the membrane cells of the transgenic pig, and that the hDPP-4 gene appears in various tissues and tails. This suggests that the expression vector is normally expressed in transgenic pigs. These results are anticipated to be a model animal to check the endocrine function for insulin resistance that occurs in a hDPP-4 transgenic pig and to increase its value for use as a material in newly developed medicines.

Dipeptidyl peptidase-4 (DPP-4) 저해제는 부작용이 적어 제2형 당뇨병의 2차 용법으로 널리 사용되고 있다. 우리는 human DPP-4 (hDPP-4) 유전자를 돼지 zygote의 전핵에 주입한 후, 1 세포 단계의 수정란을 외과적 방법으로 이식하였고, 마지막으로 꼬리에서 hDPP-4 유전자가 발현되는 형질 전환 돼지를 생산하는데 성공했다. 이식한 3두의 임신돈에서 16두의 자돈이 생산되었고, 이들 가운데 1두의 수컷 자돈에서 형질전환 개체가 확인되었다. Western blot과 RT-PCR 분석을 통해 hDPP-4가 형질 전환 돼지의 막 세포에서 강하게 발현되고, hDPP-4 유전자가 다양한 조직과 꼬리에서 각각 발현되고 있음을 확인하였다. 이것은 발현 벡터가 형질 전환 돼지에서 정상적으로 발현된다는 것을 시사한다. 이번 연구 결과는 인슐린 저항성에 대한 내분비 기능을 확인하는 모델 동물로, hDPP-4 형질 전환 돼지가 인슐린 저항성에 대한 내분비 기능을 확인할 수 있는 새로운 신약에 대한 검증의 소재로서 가치를 높일 것으로 기대된다.

Keywords

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