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Related Experiment Videos

Transgene manipulation in zebrafish by using recombinases.

Jie Dong1, Gary W Stuart

  • 1Department of Life Sciences, Indiana State University, Terre Haute, Indiana 47809, USA.

Methods in Cell Biology
|December 18, 2004
PubMed
Summary

Efficient genome engineering in zebrafish is advancing with Cre recombinase for site-specific transgene deletion. Combining Cre-loxP and SB transposon systems promises precise DNA modification and mobilization for future applications.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Zebrafish Research

Background:

  • Precise genome engineering is crucial for advancing biological research and biotechnology.
  • Zebrafish are a valuable model organism for genetic studies due to their rapid development and optical transparency.
  • Existing genome engineering tools have limitations in efficiency and specificity.

Purpose of the Study:

  • To validate methods for precise zebrafish genome reengineering using combined Cre-loxP and SB transposon systems.
  • To demonstrate the efficacy of Cre recombinase in site-specific transgene deletion in zebrafish.
  • To explore the potential for combining SB transposase-mediated transgene integration with Cre-loxP-mediated modification.

Main Methods:

  • Utilized Cre recombinase to mediate site-specific deletion of transgenes in zebrafish.

Related Experiment Videos

  • Delivered Cre recombinase via microinjection of mRNA or plasmid DNA.
  • Assessed target site utilization efficiency for transient and integrated transgenes.

    • Main Results:

    • Cre recombinase demonstrated effective site-specific deletion of transgenes in zebrafish.
    • Target site utilization efficiency approached 100%, irrespective of transgene integration status.
    • Microinjection of Cre mRNA showed slightly higher efficiency than plasmid DNA.

    Conclusions:

    • Cre recombinase can efficiently mediate site-specific transgene deletion in zebrafish.
    • A combined Cre-loxP and SB transposon system holds promise for precise zebrafish genome engineering.
    • This system offers potential for deleting, replacing, or mobilizing large DNA fragments for various applications, including gene therapy and stock improvement.