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Embryo Microinjection Techniques for Efficient Site-Specific Mutagenesis in Culex quinquefasciatus
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Genetics
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JoVE Journal Genetics
Embryo Microinjection Techniques for Efficient Site-Specific Mutagenesis in Culex quinquefasciatus

Embryo Microinjection Techniques for Efficient Site-Specific Mutagenesis in Culex quinquefasciatus

DOI:
10.3791/61375-v

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05:59 min

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May 24, 2020

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, , , ,
1Section of Cell and Developmental Biology,University of California, San Diego, 2Department of Entomology and Plant Patholog,Auburn University, 3Tata Institute for Genetics and Society,University of California, San Diego

Chapters

  • 00:04Introduction
  • 00:51Alignment of C. quinquefasciatus Pre-blastoderm Stage Embryos
  • 01:46Embryo Microinjection
  • 03:32Embryo Recovery and Hatching
  • 04:08Results: Survival and Mutation Rates of Embryos Injected with Single and Multiplexed gRNAs Targeting White
  • 05:25Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes microinjection procedures for Culex quinquefasciatus embryos that are optimized to work with CRISPR/Cas9 gene editing tools. This technique can efficiently generate site-specific, heritable, germline mutations that can be used for building genetic technologies in this understudied disease vector.

Tags

Embryo MicroinjectionCulex QuinquefasciatusSite-specific MutagenesisEgg Raft SeparationHalocarbon OilNeedle PullingMicroinjection ProtocolGenetic ToolsVector ControlGene Function Study

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