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CRISPR-Cas9 Genome Editing of Rat Embryos using Adeno-Associated Virus (AAV) and 2-Cell Embryo Electroporation
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JoVE Journal Genetics
CRISPR-Cas9 Genome Editing of Rat Embryos using Adeno-Associated Virus (AAV) and 2-Cell Embryo Electroporation

CRISPR-Cas9 Genome Editing of Rat Embryos using Adeno-Associated Virus (AAV) and 2-Cell Embryo Electroporation

DOI:
10.3791/66069-v

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08:24 min

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March 15, 2024

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, , , , ,
1Animal Modeling Core,University of Missouri, 2Comparative Medicine Program,University of Missouri, 3Rat Resource and Research Center,University of Missouri, 4Department of Veterinary Pathobiology,University of Missouri

Chapters

  • 00:05Introduction
  • 01:13AAV-mediated DNA repair template delivery
  • 02:54Electroporation preparation
  • 03:432-cell embryo electroporation
  • 06:19Representative results
  • 07:48Conclusion

Summary

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This protocol presents an optimized approach for producing genetically modified rat models. Adeno-associated virus (AAV) is used to deliver a DNA repair template, and electroporation is used to deliver CRISPR-Cas9 reagents to complete the genome editing process in 2-cell embryo.

Tags

CRISPR-Cas9Genome EditingRat EmbryosAdeno-associated Virus (AAV)2-cell Embryo ElectroporationHomology Directed RepairKnock-in Rat ModelsDNA Repair TemplateDNA InsertionDNA SubstitutionViral TransductionCRISPR RNP ComplexesElectroporation

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