Liposome-Mediated Electric Gene Delivery into Fetal and Adult Gonads

Liposome을 매개로 한 태아 및 웅성 생식선으로의 전기적 유전자 도입

  • Choi, S. C. (School of Life Sciences and Biotechnology, Korea University) ;
  • S. K. Choi (School of Life Sciences and Biotechnology, Korea University) ;
  • S. S. Choi (School of Life Sciences and Biotechnology, Korea University) ;
  • S. U. Kim (School of Life Sciences and Biotechnology, Korea University) ;
  • N. N. Cho (School of Life Sciences and Biotechnology, Korea University) ;
  • J. Y. Jung (School of Life Sciences and Biotechnology, Korea University) ;
  • C. S. Park (Division of Animal Science & Resources, Research Center for Transgenic Cloned Pigs, Chungnam National Universit) ;
  • S. H. Lee (Kongju University) ;
  • S. H. Lee (School of Life Sciences and Biotechnology, Korea University)
  • Published : 2004.03.01

Abstract

Gene delivery is one of the keen interests in animal industry as well as research on gene functions. Some of the in vivo gene delivery techniques have been successively used in various tissues for the gene therapy and transgenesis. Despite intensive efforts, it still remains to overcome problems of limited local and regional administration and low transgene expression. To improve the efficiency of gene delivery, a new procedure was tested. We injected exogenous DNA containing LacZ into the female or male gonads and then pulsed electric field. Electroporated gonads showed positive X-gal staining in many seminiferous tubules of the porcine fetal gonads. Exogenously introduced LacZ genes were also expressed in female porcine gonad. In addition, we demonstrated efficient gene delivery in gonad of adult mouse. Furthermore, we succeed to generate genetically modified germline cells showing GFP and positive X-gal signals. The results suggest that the newly developed gene delivery is an effective way of in vivo transfection in mammals. The developed gene delivery procedure should be useful in producing transgenic animals when combined with primary cell culture and nuclear transplantation.

Keywords

References

  1. Chemomordik, L. V., Papahadjopoulos, D. and Tsong, T. Y. 1991. Increased binding of liposomes to cells by electric treatment. Biochim. Biophys. Acta 1070:193-197 https://doi.org/10.1016/0005-2736(91)90163-3
  2. Clark, P. R. and Hersh, E. M. 1999. Cationic lipid-mediated gene transfer: current concepts. Curro Opin. Mol. Ther. 1:158-176
  3. Eide, F. F., Eisenberg, S. R. and Sanders, T. A. 2000. Electroporation-mediated gene transfer in free-swimming embryonic Xenopus laevis. FEBS Lett. 486:29-32 https://doi.org/10.1016/S0014-5793(00)02124-4
  4. Hara, T., Tan, Y. and Huang, L. 1997. In vivo gene delivery to the liver using reconstituted chylomicron remnants as a novel nonviral vector. Proc. Natl. Acad. Sci. USA. 94: 14547-14552
  5. Herweijer, H. and Wolff, J. A. 2003. Progress and prospects: naked DNA gene transfer and therapy. Gene Ther. 10: 453-458
  6. Inoue, T. and Krumlauf, R. 2001. An impulse to the brain-using in vivo electroporation. Nat. Neurosci. 4: 1156-1158
  7. Kikuchi, N., Nakamura, S., Ohtsuka, M., Kimura, M. and Sato, M. 2002. Possible mechanism of gene transfer into early to mid-gestational mouse fetuses by tail vein injection. Gene Ther. 9:1529-1541
  8. Kootstra, N. A. and Verma, I. M. 2003. Gene therapy with viral vectors. Annu. Rev. Phannacol. Toxicol. 43:413-439 https://doi.org/10.1146/annurev.pharmtox.43.100901.140257
  9. Liu, F. and Huang, L. 2002. Electric gene transfer to the liver following systemic administration of plasmid DNA. Gene Ther. 9:1116-1119
  10. Matsuura, M., Yamazaki, Y., Sugiyama, M., Kondo, M., Ori, H., Nango, M. and Oku, N. 2003. Polycation liposome-mediated gene transfer in vivo. Biochim. Biophys. Acta 1612:136-143
  11. Momose, T., Tonegawa, A., Takeuchi, J., Ogawa, H., Umesono, K. and Yasuda, K. 1999. Efficient targeting of gene expression in chick embryos by microelectroporation. Dev. Growth Differ. 41:335-344 https://doi.org/10.1046/j.1440-169X.1999.413437.x
  12. Muramatsu, T., Nakamura, A. and Park, H. M. 1998. In vivo electroporation: a powerful and convenient means of nonviral gene transfer to tissues of living animals. Int. J. Mol. Med. 1:55-62
  13. Ogura, T. 2002, In vivo electroporation: a new frontier for gene delivery and embryology. Differentiation 70:163-171
  14. Robinson, H. P. 1973. Sonar measurement of fetal crownrump length as means of assessing maturity in first trimester of pregnancy. Hr. Med. J. 4:28-31 https://doi.org/10.1136/bmj.4.5883.28
  15. Verma, I. M. and Somia, N. 1997. Gene therapy - promises, problems and prospects. Nature 389:239-242(Received 12 February 2004 / Accepted 15 March 2004)