Transgenic Rabbit Service

Introduction to transgenic rabbit production

To use rabbit as a research and development tool for your project, it is necessary to generate transgenic rabbits, in most if not all cases.

In theory, there are three major approaches to produce a transgenic rabbit line: 1) pronuclear DNA microinjection; 2) embryonic stem cell; and 3) somatic cell nuclear transfer. Table 2 compares these three approaches.

DNA microinjection: in 1980, Gordon et al.¹ established the first transgenic mice by pronuclear microinjection and since then, numerous transgenic mouse lines have come into existence each year.  Briefly, DNA construct is injected to the pronuclear of a zygote stage embryos, via a fine pulled needled operated by a micromanipulation system. By using this technology, larger transgenic mammals, including rats, rabbits, sheep, pigs and cattle ² have also been made. It is the primary method for the production of transgenic rabbits.

For more information:

USDA          Wiki        ILAR journal

ES cells: ES cell technology has developed dramatically in the past decade. Stable ES cell lines have been established in many mammalian species, including hamster¹⁰, mink¹¹, pigs¹², sheep¹², cattle¹³ and human⁴. In mice, genetic manipulation can be applied to ES cells. After confirming the genotype (e.g., knockout), the ES cells are injected to a blastocyst stage embryo. If success, chimeric mice are born carrying the transgenic genotype germline cells. After breeding, the desirable transgenic mice will be generated. Unfortunately, despite all the achievements and efforts, only murine ES cells have successfully transmitted the ES cell genome through the germline. In other words, we can only produce gene targeted transgenic animals in mouse, using the ES cell method. We are not yet able to produce transgenic rabbits via ES cells.

For more information,

Wiki            Knockout Science

Somatic cell nuclear transfer: animal cloning via somatic cell nuclear transfer offers a technology for multiplying in large numbers genetically valuable animals.  Particularly to our interest, somatic cell cloning (not stem cell or embryonic blastomeres cloning) provides a new route to produce gene targeted transgenic animals.  Since the birth of Dolly ³, cloned animals from adult tissues have been successfully reported in cattle ⁴, pigs ⁵, goats ⁶, mice ⁷, rabbits ⁸, cats⁹, horses ¹⁰, mules ¹¹, rats ¹² and dogs ¹³.  After gene targeting in cultured somatic cells, such cells can be used for SCNT, and gene targeted transgenic animals can be born via this approach. Already, numerous companies and institutes worldwide have incorporated cloning technology into their transgenic animals programs to produce cloned/transgenic animals for pharmaceutical (e.g., milk recombinant proteins) or medical applications (e.g., xenotransplantation for pig-to-human organ transplantation).  The first successful case of rabbit cloning ⁸ was reported in April 2002.  Unfortunately, to date, still no group in the world is able to use a gene targeted transgenic somatic cell to produce live rabbit clones. It is still in the stage of research.

For more information,

Wiki      Research Defence Society, UK      Animal Science, UC Davis

References

¹           J. W. Gordon, G. A. Scangos, D. J. Plotkin et al., Proc Natl Acad Sci U S A 77 (12), 7380 (1980).

²           J. Fan and T. Watanabe, Pharmacol Ther 99 (3), 261 (2003).

³           I. Wilmut, A. E. Schnieke, J. McWhir et al., Nature 385 (6619), 810 (1997).

⁴           C. Kubota, H. Yamakuchi, J. Todoroki et al., Proc Natl Acad Sci U S A 97 (3), 990 (2000); J. B. Cibelli, S. L. Stice, P. J. Golueke et al., Science 280 (5367), 1256 (1998); Y. Kato, T. Tani, Y. Sotomaru et al., Science 282 (5396), 2095 (1998); D. N. Wells, P. M. Misica, and H. R. Tervit, Biol Reprod 60 (4), 996 (1999); J. R. Hill, Q. A. Winger, C. R. Long et al., Biol Reprod 62 (5), 1135 (2000).

⁵           J. Betthauser, E. Forsberg, M. Augenstein et al., Nat Biotechnol 18 (10), 1055 (2000); A. Onishi, M. Iwamoto, T. Akita et al., Science 289 (5482), 1188 (2000); I. A. Polejaeva, S. H. Chen, T. D. Vaught et al., Nature 407 (6800), 86 (2000).

⁶           A. Baguisi, E. Behboodi, D. T. Melican et al., Nat Biotechnol 17 (5), 456 (1999); X. Zou, Y. Chen, Y. Wang et al., Cloning 3 (1), 31 (2001); C. L. Keefer, R. Keyston, A. Lazaris et al., Biol Reprod 66 (1), 199 (2002).

⁷           T. Wakayama, A. C. Perry, M. Zuccotti et al., Nature 394 (6691), 369 (1998).

⁸           P. Chesne, P. G. Adenot, C. Viglietta et al., Nat Biotechnol 20 (4), 366 (2002).

⁹           T. Shin, D. Kraemer, J. Pryor et al., Nature 415 (6874), 859 (2002).

¹⁰          C. Galli, I. Lagutina, G. Crotti et al., Nature 424 (6949), 635 (2003).

¹¹          G. L. Woods, K. L. White, D. K. Vanderwall et al., Science 301 (5636), 1063 (2003).

¹²          Q. Zhou, J. P. Renard, G. Le Friec et al., Science 302 (5648), 1179 (2003).

¹³          B. C. Lee, M. K. Kim, G. Jang et al., Nature 436 (7051), 641 (2005).