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형질전환 마우스 생산 및 표현형에 pronuclear microinjection 이 미치는 영향 연구

A Potential Demerit of the Pronuclear Microinjection Technique

  • 왕애국 (한국생명공학연구원 유전체연구센터) ;
  • 김선욱 (한국생명공학연구원 유전체연구센터) ;
  • 문형배 (원광대학교, 의과대학, 병리학교실) ;
  • 현병화 (국가영장류센터) ;
  • 남기환 (바이오 평가센터) ;
  • 서준교 (한림대학교, 의과대학, 실험동물센터) ;
  • 김남순 (한국생명공학연구원 유전체연구센터) ;
  • 유대열 (한국생명공학연구원 유전체연구센터) ;
  • 이동석 (한국생명공학연구원 유전체연구센터)
  • Wang, Ai-Guo (Laboratory of Human Genomics, Genetic Resources Center) ;
  • Kim, Sun-Uk (Laboratory of Human Genomics, Genetic Resources Center) ;
  • Moon, Hyung-Bae (Department of Pathology, School of Medicine and Institute of Medical Science, Wonkwang University) ;
  • Hyun, Byung-Hwa (Korea National Primate Research Center) ;
  • Nam, Ki-Hoan (Lab of Animal Model Evaluation, Korea Research Institute of Bioscience and Biotechnology(KRIBB)) ;
  • Suh, Jun-Gyo (Experimental Animal Center, College of Medicine, Hallym University) ;
  • Kim, Nam-Soon (Laboratory of Human Genomics, Genetic Resources Center) ;
  • Yu, Dae-Yeul (Laboratory of Human Genomics, Genetic Resources Center) ;
  • Lee, Dong-Seok (Laboratory of Human Genomics, Genetic Resources Center)
  • 발행 : 2006.07.31

초록

현재, 유전자의 in vivo 기능을 연구하기 위해 가장 많이 이용되고 있는 transgenic mice를 생산하기 위한 기본적으로 이용되고 있는 방법이 one cell-stage embryo에 pronuclear microinjection (PMI)이다. 그러나, 이 PMI 후에 one cell-stage embryo들의 생존율은 현저히 감소 (65.4%)할 뿐만 아니라 PMI 후의 embryo의 출생률(26.4%)이 PMI 처리를 하지 않은 것 (41.9%) 보다 현저히 낮다. 더욱이, PMI 방법에 의해 태어난 transgenic founder들의 간 조직에 병리학적 변화가 14.8% 정도에 대해서 같은 한배의 새끼 non-transgenic founder들의 경우도 간 조직에 병리학적 변화가 14.3%로 나타났다. 결론적으로, 이 PMI 방법에 의한 염색체에 물리적 손상은 형질전환 마우스의 생산 및 표현형에 영향을 미치는 잠재적 요소로 생각된다.

Pronuclear microinjection (PMI) is a primary method for producing transgenic mice and offers a powerful tool for investigating gene function in vivo. The method has several reported advantages and disadvantages. Here, we report another potential shortcoming. The survival rate of fertilized one cell-stage embryos was significantly reduced after PMI procedure (65.4% (1202/1838)). In addition, the proportion of embryos developing to full-term was also significantly lower than that of embryos not undergoing PMI (26.5% (319/1202) vs 41.9% (57/136)). Moreover, 3 out of 21 (14.3%) founder control mice which were non-transgene-carrying littermates of transgenic founders showed histopathological changes in their liver, which was comparable to that in of transgenic lineages (4 out of 27 (14.8%)). In conclusion, the mechanical damages in chromosomes occurring during PMI procedure may be a potential factor influencing phenotypes of transgenic mice.

키워드

참고문헌

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