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Next Generation Transgenic Rat Model Production.

Wanda E Filipiak1, Elizabeth D Hughes1, Galina B Gavrilina1

  • 1Transgenic Animal Model Core, University of Michigan Medical School, Ann Arbor, MI, USA.

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Summary
This summary is machine-generated.

This study details advanced methods for creating genetically engineered rat models using CRISPR/Cas9 technology. Precise genome editing via microinjection enables the development of superior research models for biomedical applications.

Keywords:
CRISPR/Cas9DNAGene editingGene knockout techniquesTransgenesTransgenic rats

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Area of Science:

  • Genetics
  • Molecular Biology
  • Animal Models

Background:

  • CRISPR/Cas9 technology significantly enhances the efficiency of modifying rat genomes.
  • Traditional microinjection methods often lead to random, multicopy transgene integration.
  • Best practices advocate for precise, single-copy transgene integration for improved model design.

Purpose of the Study:

  • To describe the next generation of genetically engineered rat models created by microinjection.
  • To highlight the use of homology-directed repair for precise genome modification.
  • To establish best practices for generating advanced transgenic rat models.

Main Methods:

  • Utilizing CRISPR/Cas9 for targeted genome modification in rat zygotes.
  • Employing pronuclear microinjection to deliver genome editing reagents.
  • Surgical transfer of microinjected zygotes to pseudopregnant rats to generate G0 founder animals.

Main Results:

  • Achieved precise, single-copy transgene integration through homology-directed repair.
  • Successfully generated genetically modified G0 founder rats carrying novel genetic modifications.
  • Demonstrated the expression of reporter proteins (e.g., eGFP, Cre recombinase) in new rat models.

Conclusions:

  • Pronuclear microinjection is the most effective method for delivering genome modification reagents for creating precise transgenic rat models.
  • These advanced rat models are crucial for accelerating biomedical research.
  • Successful generation relies on obtaining high-quality zygotes, reagent purification, and efficient embryo transfer.