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Related Concept Videos

Transgenic Organisms00:53

Transgenic Organisms

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Related Experiment Video

Updated: Aug 8, 2025

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation
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Published on: August 21, 2010

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Xenopus Transgenesis Using the pGateway System.

Liliya Nazlamova1

  • 1Clinical and Experimental Sciences, South Academic Block, Southampton General Hospital, Southampton, UK. l.nazlamova@soton.ac.uk.

Methods in Molecular Biology (Clifton, N.J.)
|February 28, 2023
PubMed
Summary

This study details a method for creating heritable genome modifications in eukaryotic organisms using I-SceI. The process involves engineering a plasmid construct and integrating it into Xenopus embryos for real-time gene expression studies.

Keywords:
Plasmid cloningSite-specific recombinationTransgenesisXenopus

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Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Genetics

Background:

  • Transgenic technologies enable the study of gene function and regulation in vivo.
  • Heritable genome modifications are crucial for understanding developmental processes and gene expression patterns.
  • The I-SceI system offers a powerful tool for precise genome engineering in various eukaryotic systems.

Purpose of the Study:

  • To describe a protocol for generating transgenic Xenopus embryos using the I-SceI system.
  • To outline the construction of a modular plasmid using the pTransgenesis system.
  • To detail the integration of engineered DNA constructs into the Xenopus genome for studying gene expression.

Main Methods:

  • Engineering of a modular plasmid construct (pTransgenesis) containing specific DNA sequence modules.
  • Co-injection of the plasmid construct and I-SceI endonuclease into Xenopus embryos.
  • Screening of injected embryos to identify and confirm successful transgenic integration.

Main Results:

  • Successful generation of transgenic Xenopus embryos with heritable genome modifications.
  • Demonstration of the pTransgenesis system for assembling complex DNA constructs.
  • Validation of the I-SceI-mediated integration method for creating stable transgenic lines.

Conclusions:

  • The described transgenic approach using I-SceI and pTransgenesis is effective for creating heritable modifications in Xenopus.
  • This method facilitates real-time, in vivo studies of gene expression patterns.
  • The protocol provides a valuable tool for genetic research in developmental biology.