CRISPR/Cas9 gene disruption studies in F0 Xenopus tadpoles: understanding development and disease in the frog

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract

CRISPR/Cas9 has become the favorite method for gene knockouts in a range of vertebrate model organisms due to its ease of use and versatility. Gene-specific guide RNAs can be designed to a unique genomic sequence and used to target the Cas9 endonuclease, which causes a double-stranded break at the desired locus. Repair of the breaks through non-homologous end joining often results in the deletion or insertion of several nucleotides, which frequently result in nonsense mutations. Xenopus frogs have long been an excellent model organism in which to study gene function, and they have proven to be useful in gene-editing experiments, especially the diploid species, X. tropicalis. In this chapter, we present our protocols for gene disruption in Xenopus, which we regularly use to investigate developmental processes and model human genetic disease.
Original languageEnglish
Title of host publicationDNA Manipulation and Analysis
EditorsGarry Scarlett
Place of PublicationNew York
PublisherHumana Press
Chapter10
Pages111-130
Number of pages20
Edition1st
ISBN (Electronic)9781071630044
ISBN (Print)9781071630037
DOIs
Publication statusPublished - 1 Mar 2023

Publication series

NameMethods in Molecular Biology
Volume2633
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • CRISPR/Cas9
  • Xenopus tropicalis
  • Xenopus laevis
  • gene knockout
  • microCT

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