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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

431
The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
431

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Related Experiment Video

Updated: Sep 20, 2025

Primordial Germ Cell Transplantation for CRISPR/Cas9-based Leapfrogging in Xenopus
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CRISPR/Cas9-based simple transgenesis in Xenopus laevis.

Yuki Shibata1, Miyuki Suzuki2, Nao Hirose3

  • 1Center for the Development of New Model Organisms, National Institute for Basic Biology, National Institutes of Natural Sciences, Okazaki, Aichi, Japan.

Developmental Biology
|June 11, 2022
PubMed
Summary

Researchers developed a new CRISPR/Cas9 method for targeted gene insertion in Xenopus laevis frogs. This technique enables efficient generation of transgenic animals for studying gene function and reporter assays.

Keywords:
CRISPR/Cas9Target integrationTransgenesisXenopus laevistgfbr2like

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Manipulation and In Vitro Maturation of Xenopus laevis Oocytes, Followed by Intracytoplasmic Sperm Injection, to Study Embryonic Development
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Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation
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Related Experiment Videos

Last Updated: Sep 20, 2025

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Manipulation and In Vitro Maturation of Xenopus laevis Oocytes, Followed by Intracytoplasmic Sperm Injection, to Study Embryonic Development
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Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation
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Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation

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Area of Science:

  • Developmental Biology
  • Genetics
  • Molecular Biology

Background:

  • Transgenic techniques are crucial for understanding gene regulation and function.
  • Existing Xenopus transgenic methods involve random DNA integration.
  • Random integration limits precise genetic manipulation and analysis in Xenopus.

Purpose of the Study:

  • To establish a simple and efficient targeted transgenesis technique in Xenopus laevis.
  • To overcome the limitations of random integration in current transgenic methods.
  • To facilitate reporter assays and gene function analysis in Xenopus.

Main Methods:

  • Developed a CRISPR/Cas9-based targeted transgenesis method.
  • Utilized Cas9 ribonucleoprotein (RNP) targeting the tgfbr2l locus as a harbor site.
  • Co-injected RNP and a donor plasmid into one-cell stage Xenopus laevis embryos.

Main Results:

  • Achieved approximately 10% faithful reporter gene expression in F0 generation (crispants).
  • Demonstrated efficient germline transmission of transgenes.
  • Observed stable transgene expression in F1 offspring in a promoter/enhancer-specific manner.

Conclusions:

  • The new CRISPR/Cas9 method enables simple and targeted transgenesis in Xenopus laevis.
  • This technique facilitates the generation of transgenic Xenopus with stable transgene expression.
  • The method is expected to become a valuable tool for gene function studies and reporter assays in Xenopus.