Efficient Transgenesis in African Turquoise Killifish Using a Gibson Assembly-Based Tol2 Transposon System

Area of Science:

• * Genetics and Molecular Biology
• * Vertebrate Aging Research
• * Regenerative Medicine

Background:

• * The African killifish (Nothobranchius furzeri) is a valuable genetic model for studying vertebrate aging and regeneration due to its short lifespan.
• * Genetically modified organisms are crucial for investigating the molecular underpinnings of biological processes.
• * Existing methods for generating transgenic killifish can be inefficient, limiting their use in research.

Purpose of the Study:

• * To establish a highly efficient protocol for generating transgenic African killifish.
• * To develop a streamlined pipeline for creating genetically modified killifish for research purposes.
• * To facilitate gene expression studies and manipulations in Nothobranchius furzeri.

Main Methods:

• * Utilized the Tol2 transposon system for random genomic integration of transgenes.
• * Employed Gibson assembly for rapid construction of transgenic vectors.
• * Incorporated an eye-specific marker for efficient identification of transgenic individuals.

Main Results:

• * Developed a protocol demonstrating high efficiency in generating transgenic African killifish.
• * Successfully created transgenic vectors with desired gene-expression cassettes and identification markers.
• * The pipeline allows for quick assembly and generation of genetically modified fish.

Conclusions:

• * The new protocol significantly enhances the ability to generate transgenic African killifish.
• * This advancement provides a powerful tool for transgenic reporter assays and gene-expression studies.
• * The developed pipeline will accelerate research in aging and regeneration using Nothobranchius furzeri.