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Production of Transgenic Rodents by the Microinjection of Cloned DNA into Fertilized One-Celled Eggs

  • David Murphy
Protocol
Part of the METHODS IN MOLECULAR BIOLOGY™ book series (MIMB, volume 461)

1. Introduction

A pioneering experiment in the early 1980s demonstrated that microinjection of a recombinant growth hormone gene into the pronuclei of fertilized one-celled mouse embryos resulted in inheritable changes in the growth of the resulting “transgenic” mice and their offspring (1). Mammalian transgenic experiments have since contributed tremendously to our understanding of numerous com plex biological processes. The power of the technique lies in that it allows the function, and the developmental and physiological regulation, of almost any gene or protein to be studied within the context of the normal processes occur ring in the whole animal.

How DNA integrates into the host chromosome is not known, but some insights can be inferred from a study made on the state and organization of the inserts found in transgenic mice (2). Approximately 70% of the mice carry exogenous DNA in all their somatic and germ cells, implying that integration took place prior to the first round of...

Keywords

Body Wall Disposable Syringe Foster Mother Surrogate Mother Injection Chamber 
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References

  1. 1.
    Palmiter RD, Brinster RL, Hammer RE, Trumbauer ME, Rosenfeld MG, Birberg NC et al. (1982) Dramatic growth of mice that develop from eggs microinjected with metallothioneine-growth hormone fusion genes. Nature 300:611–615.CrossRefPubMedGoogle Scholar
  2. 2.
    Palmiter RD, Brinster RL (1986) Germ line transformation of mice. Ann Rev Genet 20:465–499.CrossRefPubMedGoogle Scholar
  3. 3.
    Lacy E, Roberts S, Evans EP, Burtenshaw MD, Constantini F (1983) A foreign beta globin gene in transgenic mice—integration at abnormal chromosomal positions and expression in inappropriate tissues. Cell 34:343–358.CrossRefPubMedGoogle Scholar
  4. 4.
    Krumlauf R, Chapman VM, Hamme, RE, Brinster RL, Tilghman SM (1985) Differential expression of α-fetoprotein genes on the inactive X chromosome in extra-embryonic and somatic tissues of a transgenic rodent line. Nature 319:224–226.CrossRefGoogle Scholar
  5. 5.
    Wells T, Carter DA (2001) Genetic engineering of neural function in transgenic rodents: towards a comprehensive strategy? J Neurosci Methods 108:111–130.CrossRefPubMedGoogle Scholar
  6. 6.
    Boyton RJ, Altmann DM (2002) Transgenic models of autoimmune disease. Clin Exp Immunol 127:4–11.CrossRefPubMedGoogle Scholar
  7. 7.
    Wu X, Pandolfi PP (2001) Mouse models for multistep tumorigenesis. Trends Cell Biol 11:S2–S9.PubMedGoogle Scholar
  8. 8.
    Waller SJ, Ho M, Murphy D (1995) In: Glover DM, Hames BD (eds) DNA Cloning 4: a practical approach IRL, New York, pp. 185–229.Google Scholar
  9. 9.
  10. 10.
    Brinster RL, Chen HY, Trunbauer ME, Yagle MK, Palmiter RD (1985) Factors affecting the efficiency of introducing foreign DNA into mice by microinjecting eggs. Proc Natl Acad Sci USA 82:4438–4442.CrossRefPubMedGoogle Scholar
  11. 11.
    Auerbach AB, Norinsky R, Ho W, Losos K, Guo Q, Chatterjee S, Joyner AL (2003) Strain-dependent differences in the efficiency of transgenic mouse production. Transgenic Research 12:59–69.CrossRefPubMedGoogle Scholar
  12. 12.
    Vogelstein B, Gillespie D (1979) Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci USA 82:615–619.CrossRefGoogle Scholar
  13. 13.
    Constantini F, Lacy E (1981) Introduction of a rabbit B-globin gene into the mouse germ line. Nature 294:92–94.CrossRefGoogle Scholar
  14. 14.
    Leibo SP (1984) A one-step method for direct nonsurgical transfer of frozen-thawed bovine embryos. Theriogenology 21:767–790.CrossRefPubMedGoogle Scholar
  15. 15.
    Pfaff RT, Agca Y, Liu J, Woods EJ, Peter AT, Critser JK (2000) Cryobiology of rat embryos, I: determination of zygote membrane permeability coefficients for water and cryoprotectants, their activation energies, and the development of improved cryopreservation methods. Biol Reprod 63:1294–1302.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science + Business Media, LLC 2008

Authors and Affiliations

  • David Murphy
    • 1
  1. 1.Laboratories of Integrative Neurosciences and EndocrinologyUniversity of BristolBristol

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