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Related Concept Videos

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The microinjection of mouse oocytes is commonly used for both classic transgenesis (i.e., the random integration of transgenes) and CRISPR-mediated gene targeting. This protocol reviews the latest developments in microinjection, with a particular emphasis on quality control and genotyping...
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Related Experiment Video

Updated: Jan 19, 2026

Generation of Genetically Modified Mice through the Microinjection of Oocytes
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Breeding Strategies for Genetically Modified Mice.

Jan Parker-Thornburg1

  • 1Genetically Engineered Mouse Facility, Department of Genetics, MD Anderson Cancer Center, Houston, TX, USA. jpthorn@mdanderson.org.

Methods in Molecular Biology (Clifton, N.J.)
|September 13, 2019
PubMed
Summary

Establishing genetically modified mouse lines requires specific breeding strategies tailored to the generation method. Methods like embryonic stem cell targeting and CRISPR/Cas9 gene editing necessitate germline transmission testing for successful line establishment.

Keywords:
CRISPRCRISPR/Cas9ChimeraGerm line transmissionKnock-in miceKnock-out miceMosaicTransgenic

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Area of Science:

  • Genetics
  • Animal Models
  • Molecular Biology

Background:

  • Genetically modified mice are essential research tools.
  • Various methods exist for generating genetically modified mice.
  • Each method presents unique challenges for establishing stable mouse lines.

Purpose of the Study:

  • To outline the necessary breeding strategies for establishing genetically modified mouse lines.
  • To highlight method-specific considerations for genetic modification in mice.
  • To emphasize the importance of germline transmission testing.

Main Methods:

  • Review of established techniques for generating genetically modified mice.
  • Analysis of breeding schemes for transgenic founder mice.
  • Description of germline transmission testing protocols for embryonic stem cell and CRISPR/Cas9 targeted mice.

Main Results:

  • Transgenic founder mice require outcrossing to establish lines.
  • Embryonic stem cell-targeted mice need germline transmission confirmation.
  • CRISPR/Cas9-edited mice often exhibit mosaicism, requiring germline transmission assessment.

Conclusions:

  • Successful establishment of genetically modified mouse lines depends on appropriate breeding strategies.
  • Germline transmission testing is a critical step for methods like embryonic stem cell targeting and CRISPR/Cas9 gene editing.
  • Tailored approaches are necessary for each genetic modification technique to ensure reliable mouse line development.