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Related Experiment Videos

Efficient metaphase II transgenesis with different transgene archetypes.

A C Perry1, A Rothman, J I de las Heras

  • 1The Rockefeller University, 1230 York Avenue, New York, NY 10021, USA. tperry@advancedcell.com

Nature Biotechnology
|November 2, 2001
PubMed
Summary

Mouse oocytes can efficiently create transgenic animals using large DNA constructs, including artificial chromosomes. This method achieves high efficiency and reliable transmission of transgenes to offspring.

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Area of Science:

  • Genomics and Biotechnology
  • Mammalian Transgenesis
  • Reproductive Biology

Background:

  • Producing transgenic animals requires mobilizing large DNA segments for genome characterization and biotechnology.
  • Previous work demonstrated efficient transgenesis using small transgenes (<5 kilobases) in mouse metaphase II (mII) oocytes.

Purpose of the Study:

  • To extend the application of mII oocyte transgenesis to larger DNA constructs.
  • To assess the efficiency and germline transmission of large transgenes using mII transgenesis.

Main Methods:

  • Coinjection of sperm with large transgenes (11.9-170 kilobases), including bacterial artificial chromosomes (BACs) and mammalian artificial chromosomes (MACs), into mouse mII oocytes.
  • Analysis of offspring for transgene integration and transmission.

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Main Results:

  • mII transgenesis successfully incorporated larger transgenes, including BAC and MAC constructs.
  • Transgenesis efficiency for large constructs ranged from 11-47% of offspring.
  • Over 95% of transgenic founders efficiently transmitted the large transgenes to their progeny.

Conclusions:

  • Mouse mII oocyte transgenesis is a robust method for efficiently delivering large DNA constructs.
  • This technique significantly advances the ability to generate transgenic animals with complex genetic material.