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Generating Genetically Engineered Mice Using a Spermatogonial Stem Cell-Mediated Method.

Takuya Sato1, Takehiko Ogawa2,3

  • 1Laboratory of Biopharmaceutical and Regenerative Sciences, Institute of Molecular Medicine and Life Science, Yokohama City University Association of Medical Science, Yokohama, Kanagawa, Japan. tsato@yokohama-cu.ac.jp.

Methods in Molecular Biology (Clifton, N.J.)
|October 25, 2018
PubMed
Summary
This summary is machine-generated.

Mouse spermatogonial stem cells (SSCs) can be cultured and transplanted to generate sperm for producing offspring. This method enables the creation of genetically modified mice using genome editing techniques.

Keywords:
CRISPR/Cas9Cell transplantationGS cellGene targetingGenome editingIntratubular injectionSpermatogonial stem cell

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Area of Science:

  • Reproductive biology
  • Stem cell research
  • Genetics

Background:

  • Mouse spermatogonial stem cells (SSCs) possess self-renewal and differentiation capabilities.
  • Cultured SSCs, termed germline stem (GS) cells, maintain pluripotency in vitro.
  • Transplanted GS cells can restore spermatogenesis in host testes.

Purpose of the Study:

  • To describe a method for genome editing-mediated modification of GS cells.
  • To detail the transplantation of modified GS cells into mouse testes.
  • To enable the generation of genetically modified mice.

Main Methods:

  • Culture and expansion of mouse SSCs (GS cells).
  • Application of genome editing techniques to modify GS cells.
  • Intratesticular transplantation of modified GS cells into recipient mice.

Main Results:

  • Successful long-term culture and maintenance of GS cell properties.
  • Demonstration of GS cell colonization and spermatogenesis establishment in host testes.
  • Production of offspring derived from transplanted, genome-edited GS cells.

Conclusions:

  • Genome editing of GS cells followed by transplantation is a viable strategy for generating genetically modified mice.
  • This approach offers a powerful tool for genetic manipulation and functional studies in mice.
  • The method preserves the reproductive potential of modified germline stem cells.