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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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遗传学
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JoVE 杂志 遗传学
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

1,244 Views

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08:24 min

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March 15, 2024

DOI:

10.3791/66069-v

DOI:
10.3791/66069-v

•

08:24 min
March 15, 2024

•

23 Views
, , , , ,
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

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.

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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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J. Davis, D., McNew, J. F., Walls, J. N., Bethune, C. E., Oswalt, P. S., Bryda, E. C. CRISPR-Cas9 Genome Editing of Rat Embryos using Adeno-Associated Virus (AAV) and 2-Cell Embryo Electroporation. J. Vis. Exp. (205), e66069, doi:10.3791/66069 (2024).

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