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Embryo Microinjection and Knockout Mutant Identification of CRISPR/Cas9 Genome-Edited Helicoverpa Armigera (Hübner)
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JoVE Journal Behavior
Embryo Microinjection and Knockout Mutant Identification of CRISPR/Cas9 Genome-Edited Helicoverpa Armigera (Hübner)

Embryo Microinjection and Knockout Mutant Identification of CRISPR/Cas9 Genome-Edited Helicoverpa Armigera (Hübner)

DOI:
10.3791/62068-v

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06:37 min

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July 01, 2021

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, , , , ,
1School of Chemical and Environmental Engineering,China University of Mining and Technology, 2State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection,Chinese Academy of Agricultural Sciences, 3Guangdong Laboratory of Lingnan Modern Agriculture, Shenzhen; Agricultural Genomics Institute at Shenzhen,Chinese Academy of Agricultural Sciences

Chapters

  • 00:05Introduction
  • 00:29Design of Gene-Specific Primers and Preparation of sgRNA and Embryos
  • 02:18Microinjection of Embryos
  • 03:47Post-Injection Insect Rearing
  • 04:41Knock-Out Mutation Detection
  • 05:27Results: Selection and Synthesis of Target sgRNAs and Detection of Mutants by PCR and Gel Electrophoresis
  • 06:08Conclusion

Summary

Automatic Translation

Automatic Translation

Presented here is a protocol of Helicoverpa armigera (Hübner) embryo microinjection and knockout mutant identification created by CRISPR/Cas9 genome editing. Mutant insects enable further research of gene function and interaction among different genes in vivo.

Tags

Keywords: CRISPR/Cas9Genome EditingHelicoverpa ArmigeraCotton BollwormEmbryo MicroinjectionKnockout Mutant IdentificationSgRNA Target SelectionPAM SequenceEmbryo PreparationEgg CollectionMicroinjectionCas9 ProteinSgRNA

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