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Determining the Role of Maternally-Expressed Genes in Early Development with Maternal Crispants
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JoVE Journal Genetics
Determining the Role of Maternally-Expressed Genes in Early Development with Maternal Crispants

Determining the Role of Maternally-Expressed Genes in Early Development with Maternal Crispants

DOI:
10.3791/63177-v

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10:08 min

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December 21, 2021

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, ,
1Laboratory of Genetics,University of Wisconsin-Madison

Chapters

  • 00:05Introduction
  • 00:56Synthesis of Guide RNAs
  • 02:03Microinjection of CRISPR-Cas9 Cocktail into a One-Cell Zebrafish Embryo to Generate Maternal Crispants
  • 03:07Screening for Somatic INDELs in F0 Injected Embryos
  • 04:54Identification of Maternal-Effect Phenotypes in Maternal Crispant Embryos
  • 06:21Sequencing Alleles in Maternal Crispant Haploids
  • 08:18Results: Identification of Maternal Effect Phenotypes in a Rapid and Resource Efficient Manner
  • 09:08Conclusion

Summary

Automatic Translation

Automatic Translation

Early development is dependent on maternally-inherited products, and the role of many of these products is currently unknown. Herein, we described a protocol that uses CRISPR-Cas9 to identify maternal-effect phenotypes in a single generation.

Tags

Keywords: Maternal CRISPRMaternal Effect GenesEarly DevelopmentEmbryogenesisGuide RNAIn-vitro TranscriptionZebrafishEmbryo Injection

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