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Sand Fly (Phlebotomus papatasi) Embryo Microinjection for CRISPR/Cas9 Mutagenesis
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JoVE Journal Genetica
Sand Fly (Phlebotomus papatasi) Embryo Microinjection for CRISPR/Cas9 Mutagenesis

Sand Fly (Phlebotomus papatasi) Embryo Microinjection for CRISPR/Cas9 Mutagenesis

DOI:
10.3791/61924-v

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05:44 min

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November 17, 2020

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, , , , ,
1Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases,National Institutes of Health, 2University of Maryland Insect Transformation Facility,The Institute for Bioscience and Biotechnology Research

Capitoli

  • 00:04Introduction
  • 00:38Embryo Collection and Micromanipulation
  • 02:03Embryo Injections
  • 03:59Results: Representative Identification of Mosaic G0 Adults and Mutations of Interest
  • 05:07Conclusion

Summary

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This protocol details the steps of CRISPR/Cas9 targeted mutagenesis in sand flies: embryo collection, injection, insect rearing, and identification as well as selection of mutations of interest.

Tags

Keywords: Sand FlyPhlebotomus PapatasiEmbryo MicroinjectionCRISPR/Cas9MutagenesisInsect Genome EditingBlood FeedingEgg-layingFilter PaperMicroscopeInjection NeedlePneumatic Injection Controller

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