생명과학회지 (Journal of Life Science)

  • 제16권4호
  • /
  • Pages.566-570
  • /
  • 2006
  • /
  • 1225-9918(pISSN)
  • /
  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지
DOI QR코드

DOI QR Code

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
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

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

  1. Brinster, R. L., H. Y. Chen, M. E. Trumbauer, M. K. Yagle, and R. D. Palmiter. 1985. Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc. Natl. Acad. Sci. USA. 82(13), 4438-4442
  2. Frith, C. H. and J. M. Ward. 1979. A morphologic classification of proliferative and neoplastic hepatic lesions in mice. J. Environ. Pathol. Toxicol. 3(1-2), 329-351
  3. Gordon, J. W., G. A. Scangos, D. J. Plotkin, J. A. Barbosa, and F. H. Ruddle. 1980. Genetic transformation of mouse embryos by microinjection of purified DNA. Proc. Natl. Acad. Sci. USA. 77(12), 7380-7384
  4. Hogan, B., R. Beddington, F. Costantini, and E. Lacy A Laboratory Manual: Manipulating the Mouse Embryo, Second Edition edition: Cold Spring Harbor Laborary Press; 1994
  5. Maronpot, R. R., J. K. Haseman, G. A. Boorman, S. E. Eustis, G. N. Rao, and J. E. Huff. 1987. Liver lesions in B6C3F1 mice: the National Toxicology Program, experience and position. Arch. Toxicol. Suppl. 10,10-26
  6. Meng, X., H. Akutsu, K. Schoene, C. Reifsteck, E. P. Fox, S. Olson, H. Sariola, R. Yanagimachi, and M. Baetscher. 2002. Transgene insertion induced dominant male sterility and rescue of male fertility using round spermatid injection. Biol. Reprod. 66(3), 726-734 https://doi.org/10.1095/biolreprod66.3.726
  7. Nomura, T. 1997. Practical development of genetically engineered animals as human disease models. Lab. Anim. Sci. 47(2), 113-117
  8. Polites, H. G. and C. A. Pinkert Transgenic Animal Technology: A Laboratory Handbook: Academic. Press; 1994
  9. Sambrook, J., E. F. Fritsch, and T. Maniatis A Laboratory Manual: Molecular Cloning. 1989
  10. Smith, G. S., R. L. Walford, and M. R. Mickey. 1973. Lifespan and incidence of cancer and other diseases in selected long-lived inbred mice and their F 1 hybrids. J. Natl. Cancer Inst. 50(5), 1195-1213 https://doi.org/10.1093/jnci/50.5.1195
  11. Waterston, R. H., K. Lindblad-Toh, E. Birney, J. Rogers, J. F. Abril, P. Agarwal, R. Agarwala, R. Ainscough, M. Alexandersson, P. An, S. E. Antonarakis, J. Attwood, R. Baertsch, J. Bailey, K. Barlow, S. Beck, E. Berry, B. Birren, T. Bloom, P. Bork, M. Botcherby, N. Bray, M. R. Brent, D. G. Brown, S. D. Brown, C. Bult, J. Burton, J. Butler, R. D. Campbell, P. Carninci, S. Cawley, F. Chiaromonte, A. T. Chinwalla, D. M. Church, M. Clamp, C. Clee, F. S. Collins, L. L. Cook, R. R. Copley, A. Coulson, O. Couronne, J. Cuff, V. Curwen, T. Cutts, M. Daly, R. David, J. Davies, K. D. Delehaunty, J. Deri, E. T. Dermitzakis, C. Dewey, N. J. Dickens, M. Diekhans, S. Dodge, I. Dubchak, D. M. Dunn, S. R. Eddy, L. Elnitski, R. D. Emes, P. Eswara, E. Eyras, A. Felsenfeld, G. A. Fewell, P. Flicek, K. Foley, W. N. Frankel, L. A. Fulton, R. S. Fulton, T. S. Furey, D. Gage, R. A. Gibbs, G. Glusman, S. Gnerre, N. Goldman, L. Goodstadt, D. Grafham, T. A. Graves, E. D. Green, S. Gregory, R. Guigo, M. Guyer, R. C. Hardison, D. Haussler, Y. Hayashizaki, L. W. Hillier, A. Hinrichs, W. Hlavina, T. Holzer, F. Hsu, A. Hua, T. Hubbard, A. Hunt, I. Jackson, D. B. Jaffe, L. S. Johnson, M. Jones, T. A. Jones, A. Joy, M. Kamal, E. K. Karlsson, D. Karolchik, A. Kasprzyk, J. Kawai, E. Keibler, C. Kells, W. J. Kent, A. Kirby, D. L. Kolbe, I. Korf, R. S. Kucherlapati, E. J. Kulbokas, D. Kulp, T. Landers, J. P. Leger, S. Leonard, I. Letunic, R. Levine, J. Li, M. Li, C. Lloyd, S. Lucas, B. Ma, D. R. Maglott, E. R. Mardis, L. Matthews, E. Mauceli, J. H. Mayer, M. McCarthy, W. R. McCombie, S. McLaren, K. McLay, J. D. McPherson, J. Meldrim, B. Meredith, J. P. Mesirov, W. Miller, T. L. Miner, E. Mongin, K. T. Montgomery, M. Morgan, R. Mott, J. C. Mullikin, D. M. Muzny, W. E. Nash, J. O. Nelson, M. N. Nhan, R. Nicol, Z. Ning, C. Nusbaum, M. J. O'Connor, Y. Okazaki, K. Oliver, E. Overton-Larty, L. Pachter, G. Parra, K. H. Pepin, J. Peterson, P. Pevzner, R. Plumb, C. S. Pohl, A. Poliakov, T. C. Ponce, C. P. Ponting, S. Potter, M. Quail, A. Reymond, B. A. Roe, K. M. Roskin, E. M. Rubin, A. G. Rust, R. Santos, V. Sapojnikov, B. Schultz, J. Schultz, M. S. Schwartz, S. Schwartz, C. Scott, S. Seaman, S. Searle, T. Sharpe, A. Sheridan, R. Shownkeen, S. Sims, J. B. Singer, G. Slater, A. Smit, D. R. Smith, B. Spencer, A. Stabenau, N. Stange-Thomann, C. Sugnet, M. Suyama, G. Tesler, J. Thompson, D. Torrents, E. Trevaskis, J. Tromp, C. Ucla, A. Ureta-Vidal, J. P. Vinson, A. C. Von Niederhausern, C. M. Wade, M. Wall, R. J. Weber, R. B. Weiss, M. C. Wendl, A. P. West, K. Wetterstrand, R. Wheeler, S. Whelan, J. Wierzbowski, D. Willey, S. Williams, R. K. Wilson, E. Winter, K. C. Worley, D. Wyman, S. Yang, S. P. Yang, E. M. Zdobnov, M. C. Zody and E. S. Lander. 2002. Initial sequencing and comparative analysis of the mouse genome. Nature 420(6915), 520-562 https://doi.org/10.1038/nature01262
  12. Yu, D. Y., H. B. Moon, J. K. Son, S. Jeong, S. L. Yu, H. Yoon, Y. M. Han, C. S. Lee, J. S. Park, C. H. Lee, B. H. Hyun, S. Murakami, and K. K. Lee. 1999. Incidence of hepatocellular carcinoma in transgenic mice expressing the hepatitis B virus X-protein. J. Hepatol. 31(1), 123-132 https://doi.org/10.1016/S0168-8278(99)80172-X