DOI QR코드

DOI QR Code

Cock Spermatozoa Serve as the Gene Vector for Generation of ransgenic Chicken (Gallus gallus)

  • Yang, C.C. (Department of Animal Science, National Taiwan University) ;
  • Chang, H.S. (National Health Research Institutes) ;
  • Lin, C.J. (Department of Animal Science, National Taiwan University) ;
  • Hsu, C.C. (Department of Animal Science, National Taiwan University) ;
  • Cheung, J.I. (Chung Tai Institute of Health Sciences and Technology) ;
  • Hwu, L. (GeneShuttle Biopharmaceutical, Inc.) ;
  • Cheng, W.T.K. (Department of Animal Science, National Taiwan University)
  • Received : 2003.03.14
  • Accepted : 2004.01.26
  • Published : 2004.07.01

Abstract

To evaluate the feasibility of using sperm-mediated gene transfer (SMGT) for carrying foreign gene into chicken oocyte, a reporter gene, CX-EGFP, was used in this study. The reporter gene was first mixed with liposome or liposome-like compound and the mixtures were further combined with ejaculated cock spermatozoa. The spermatozoa treated with liposome and CX-EGFP mixture was subsequently coincubated with DNaseI to remove the extra DNA which insured the authenticity of positive signals. The treated sperms were then subjected to transgene (reporter gene) existence analysis and artificial insemination of laying hens. Obtained results indicated that the spermatozoa were able to take-in the foreign DNA; which was confirmed by polymerase chain reaction and Southern blot analysis. In the following experiment, fresh ejaculated sperms were mixed with CX-EGFP-liposome or CX-EGFP-liposome-like complex then used for artificial insemination of each of six laying hens. Eggs laid between day-3 and day-7 post insemination were collected. Newly hatched chicks, two out of 53 from CX-EGFP/liposome treated group and two out of 21 from CXEGFP/liposome-like treated group, were proven to be transgenic. This study suggests that SMGT is a powerful method for generating transgenic chickens.

Keywords

Transgenic Chicken;Sperm-mediated;Gene Transfer

Acknowledgement

Supported by : Council of Agriculture

References

  1. Brackett, B. G., W. Baranska, W. Sawichi and H. Koprowski. 1971. Uptake of heterologous genome by mammalian spermatozoa and its transfer to ova through fertilization. Proc. Natl. Acad. Sci. USA. 68:353-357.
  2. Patil, J. G. and H. W. Khoo. 1996. Nuclear internalization of foreign DNA by zebrafish spermatozoa and its enhancement by electroporation. J. Exp. Zool. 274:121-129.
  3. Castro, F. O., O. Hernández, C. Uliver, R. Solano, C. Milanés, A. Aguilar, A. Pérez, R. de Armas, L. Herrera and J. de la Fuente. 1991. Introduction of foreign DNA into the spermatozoa of farm animals. Theriogenology 34:1099-1110.
  4. Ebara, F. and N. Fujihara. 2000. Successful transfer of exogenously introduced gene (lacZ/MiwZ and lacZ&GFP/pkkv4-lacZ) for generations in chicken. J. Reprod. Dev. 46:177-182.
  5. Fejzo, M. S. and D. J. Slamon. 2001. Frozen Tumor Tissue Microarray Technology for Analysis of Tumor RNA, DNA, and Proteins. Am. J. Pathol. 159:1645-1650.
  6. Maione, B., C. Pittoggi, L. Achene, R. Lorenzini and C. Spadafora. 1997. Activation of endogenous nucleases in mature sperm cells upon interaction with exogenous DNA. DNA Cell Biol. 16:1087-1097.
  7. Salter, D. W., E. J. Smith, S. H. Hughes, S. E. Wright, A. M. Fadly, R. L. Witter and L. B. Crittenden. 1986. Gene insertion into the chicken germline by retroviruses. Poult. Sci. 65:1445-1458.
  8. Han, J. Y. and D. K. Jeong. 2002. Migration activity of chicken gonadal primordial germ cells (gPGCs) and post-transfer localization of LacZ-transfected gPGCs in the embryonic gonads. Asian-Aust. J. Anim. Sci. 15:1227-1231. https://doi.org/10.5713/ajas.2002.1227
  9. Ikawa, M., S. Yamada, T. Nakanishi and M. Okabe. 1999. Green fluorescent protein (GFP) as a vital marker in mammals. Curr.Top. Dev. Biol. 44:1-20.
  10. Iba, H. 2000. Gene transfer into chicken embryos by retrovirus vectors. Dev. Growth Differ. 43:213-218.
  11. Okabe, M., M. Ikawa, K. Kominami, T. Nakanishi and Y. Nishimune. 1997. ‘Green mice’ as a source of ubiquitous green cells. FEBS Lett. 407:313-319.
  12. Rottmann, O. J., R. Antes, P. H ofer and G. Maierhofer. 1992. Liposome mediated gene transfer via spermatozoa into avian egg cells, J. Anim. Breed. Genet. 109:64-70.
  13. Zoraqi, G. and C. Spadafora. 1997. Integration of foreign DNA sequences into mouse sperm sperm. DNA Cell Biol. 16:291-300.
  14. Tsai, H. J., F. S. Tseng and I. C. Liao. 1995. Electroporation of sperm to introduce foreign DNA into the genome of loach (Misgurnus anguillicaudatus). Can. T. Fish Aquatic. Sci. 52:776-787.
  15. Thoraval, P., M. Afanassieff, F. L. Cosset, F. Lasserre, G. Verdier, F. Coudert and G. Dambrine. 1995. Germline transmission of exogenous genes in chickens using helper-free ecotropic avian leukosis virus-based vectors. Transgenic Res. 4:369-376.
  16. Shemesh, M., M. Gurevich, E. Harel-Markowitz, L. Benvenisti, L. S. Shore and Y. Stram. 2000. Gene integration into bovine sperm genome and its expression in transgenic offspring. Mol. Reprod. Dev. 56:306-308.
  17. Kuznetsov, A. V., I. V. Kuznetsova and I. Y. Schit. 2000. DNA interaction with rabbit sperm cells and its transfer into ova in vitro and in vivo. Mol. Reprod. Dev. 56:292-297.
  18. Nakamura, H. and J. Funahashi. 2001. Introduction of DNA into chick embryos by in ovo electroporation. Methods 24:43-48.
  19. Ni, Y. H., H. Y. Hsu, P. J. Chen and M. H. Chang. 1999. Protamine enhances the efficiency of liposome-mediated gene transfer in a cultured human hepatoma cell line. J. Formos. Med. Assoc. 98:562-566.
  20. Koetsier, P. A., J. Schorr and W. Doerfler. 1993. A rapid optimized protocol for downward alkaline Southern blooting of DNA. Bio/Technique. 15:260-262.
  21. Longmuir, K. J., S. M. Haynes, M. E. Dickinson, J. C. Murphy, R. C. Willson and A. J. Waring. 2001. Optimization of a peptide/non-cationic lipid gene delivery system for effective microinjection into chicken embryo in vivo. Mol. Ther. 4:66-74.
  22. Nakanishi, A. and A. Iritani. 1993. Gene transfer in the chicken by sperm-mediated methods. Mol. Reprod. Dev. 36:258-261.
  23. Lavitrano, M., A. Stoppacciaro, M. L. Bacci, M. Forni, D. Fioretti, L. Pucci, C. Di Stefano, D. Lazzereschi, A. Rughetti, E. Seren, D. Alfani, R. Cortesini and L. Frati. 1999. Human decay accelerating factor transgenic pigs for enotransplantation obtained by sperm-mediated gene transfer. Transplant. Proc. 31:972-974.
  24. Lee, C. K. and J. A. Piedrahita. 2003. Transgenesis and germ cell engineering in domestic animals. Asian-Aust. J. Anim. Sci. 16:910-927.
  25. Tsai, H. J., C. H. La and H. S. Yang. 1997. Sperm as a carrier to introduce an exogenous DNA fragment into the oocyte of Japanese abalone (Haliotis divorsicolor suportexta). Transgenic Res. 6:85-95.
  26. Birchall, J. C., I. W. Kellaway and M. Gumbleton. 2000. Physical stability and in vitro gene expression efficiency of nebulised lipid-peptide-DNA complexes. Int. J. Pharm. 197:221-231.
  27. Habrova, V., M. Takac, J. Navratil, J. Macha, N. Ceskova and J. Jonak. 1996. Association of Rous sarcoma virus DNA with Xenopus laevis spermatozoa and its transfer to ova through fertilization. Mol. Reprod. Dev. 44:332-342. https://doi.org/10.1002/(SICI)1098-2795(199607)44:3<332::AID-MRD7>3.0.CO;2-N
  28. Clark, A. G. and A. Civetta. 2000. Protamine wars. Nature 403:261-262.
  29. Han, J. Y., R. N. Shoffner and K. S. Guise. 1994. Gene transfer by manipulation of primordial germ cell in the chicken. Asian-Aust. J. Anim. Sci. 7:427-434.

Cited by

  1. Use of Sperm Plasmid DNA Lipofection Combined with REMI (Restriction Enzyme-Mediated Insertion) for Production of Transgenic Chickens Expressing eGFP (Enhanced Green Fluorescent Protein) or Human Follicle-Stimulating Hormone1 vol.80, pp.5, 2009, https://doi.org/10.1095/biolreprod.108.070375
  2. Development of transgenic chicken with a gene of human granulocyte colony-stimulating factor using sperm-mediated gene transfer vol.40, pp.5, 2013, https://doi.org/10.1134/S1062359013040134
  3. Establishment of a novel, eco-friendly transgenic pig model using porcine pancreatic amylase promoter-driven fungal cellulase transgenes vol.24, pp.1, 2015, https://doi.org/10.1007/s11248-014-9817-9
  4. Sperm-mediated gene transfer: Applications and implications vol.27, pp.5, 2005, https://doi.org/10.1002/bies.20211