Increased efficiency of transgenic livestock production

Elizabeth A. Maga, R. Geoffrey Sargent, Hong Zeng, Sushma Pati, David A. Zarling, Stefanie M. Oppenheim, Nicole M B Collette, Alice L. Moyer, Janice S. Conrad-Brink, Joan D Rowe, Robert Bondurant, Gary B. Anderson, James D. Murray

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Production of transgenic livestock by pronuclear microinjection of DNA into fertilized zygotes suffers from the compounded inefficiencies of low embryo survival and low integration frequencies of the injected DNA into the genome. These inefficiencies are one of the major obstacles to the large-scale use of pronuclear microinjection techniques in livestock. We investigated exploiting the properties of recombinase proteins that allow them to bind DNA to generate transgenic animals via pronuclear microinjection. In theory, the use of recombinase proteins has the potential to generate transgenic animals with targeted changes, but in practice we found that the use of RecA recombinase-coated DNA increases the efficiency of transgenic livestock production. The use of RecA protein resulted in a significant increase in both embryo survival rates and transgene integration frequencies. Embryo survival rates were doubled in goats, and transgene integration was 11-fold higher in goats and three-fold higher in pigs when RecA protein-coated DNA was used compared with conventional DNA constructs without RecA protein coating. However, a large number of the transgenic founders generated with RecA protein-coated DNA were mosaic. The RecA protein coating of DNA is straightforward and can be applied to any species and any existing microinjection apparatus. These findings represent significant improvements on standard pronuclear microinjection methods by enabling the more efficient production of transgenic livestock.

Original languageEnglish (US)
Pages (from-to)485-496
Number of pages12
JournalTransgenic Research
Volume12
Issue number4
DOIs
StatePublished - Aug 2003

Fingerprint

Livestock
livestock production
Rec A Recombinases
genetically modified organisms
Microinjections
DNA
Recombinases
proteins
embryo (animal)
transgenic animals
Genetically Modified Animals
Embryonic Structures
livestock
Transgenes
Goats
coatings
transgenes
survival rate
goats
Zygote

Keywords

  • Efficiency
  • Livestock
  • Recombinase
  • Transgenic

ASJC Scopus subject areas

  • Genetics
  • Plant Science
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Biotechnology
  • Food Science

Cite this

Maga, E. A., Sargent, R. G., Zeng, H., Pati, S., Zarling, D. A., Oppenheim, S. M., ... Murray, J. D. (2003). Increased efficiency of transgenic livestock production. Transgenic Research, 12(4), 485-496. https://doi.org/10.1023/A:1024257906647

Increased efficiency of transgenic livestock production. / Maga, Elizabeth A.; Sargent, R. Geoffrey; Zeng, Hong; Pati, Sushma; Zarling, David A.; Oppenheim, Stefanie M.; Collette, Nicole M B; Moyer, Alice L.; Conrad-Brink, Janice S.; Rowe, Joan D; Bondurant, Robert; Anderson, Gary B.; Murray, James D.

In: Transgenic Research, Vol. 12, No. 4, 08.2003, p. 485-496.

Research output: Contribution to journalArticle

Maga, EA, Sargent, RG, Zeng, H, Pati, S, Zarling, DA, Oppenheim, SM, Collette, NMB, Moyer, AL, Conrad-Brink, JS, Rowe, JD, Bondurant, R, Anderson, GB & Murray, JD 2003, 'Increased efficiency of transgenic livestock production', Transgenic Research, vol. 12, no. 4, pp. 485-496. https://doi.org/10.1023/A:1024257906647
Maga EA, Sargent RG, Zeng H, Pati S, Zarling DA, Oppenheim SM et al. Increased efficiency of transgenic livestock production. Transgenic Research. 2003 Aug;12(4):485-496. https://doi.org/10.1023/A:1024257906647
Maga, Elizabeth A. ; Sargent, R. Geoffrey ; Zeng, Hong ; Pati, Sushma ; Zarling, David A. ; Oppenheim, Stefanie M. ; Collette, Nicole M B ; Moyer, Alice L. ; Conrad-Brink, Janice S. ; Rowe, Joan D ; Bondurant, Robert ; Anderson, Gary B. ; Murray, James D. / Increased efficiency of transgenic livestock production. In: Transgenic Research. 2003 ; Vol. 12, No. 4. pp. 485-496.
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