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Related Experiment Videos

Sperm-mediated gene transfer in mice

B Maione1, M Lavitrano, C Spadafora

  • 1Department of Experimental Medicine, Institute of General Pathology, University La Sapienza, Roma, Italy.

Molecular Reproduction and Development
|July 21, 1998
PubMed
Summary

Sperm-mediated DNA transfer can create transgenic mice, but success rates vary. A large study found a low overall efficiency, suggesting event clustering may explain inconsistent results in generating transgenic offspring.

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Area of Science:

  • Reproductive Biology
  • Genetics
  • Biotechnology

Background:

  • Sperm-mediated DNA transfer offers a simplified method for generating transgenic animals.
  • Previous studies reported controversial and variable efficiencies for this technique in mice.

Purpose of the Study:

  • To investigate the variability in sperm-mediated DNA transfer efficiency in mice.
  • To identify factors contributing to successful transgenic offspring generation.

Main Methods:

  • Sperm were incubated with plasmid DNA during capacitation.
  • Fertilization was performed with fresh mouse oocytes.
  • Cleaved embryos were transferred to pseudopregnant recipients for gestation.

Main Results:

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  • Out of 75 experiments, 13 yielded transgenic offspring (7.4% of fetuses).
  • Five experiments achieved high success rates (>85% transgenic offspring), but contributing factors remain unidentified.
  • The low frequency and clustering of successful events may explain disparate previous reports.

    • Conclusions:

    • The variability in sperm-mediated DNA transfer success in mice might be due to the clustering of rare events.
    • Identifying factors for high success could establish this as a key tool for transgenic mouse generation.
    • Further research may reveal natural mechanisms protecting against foreign DNA transfer via sperm.