Construction of irf4a Transgenic Zebrafish Using Tol2 System and Its Potential Application

Area of Science:

• Zebrafish as a model organism in biomedical research.
• Molecular biology techniques for genetic modification.
• Immunology and the role of transcription factors.

Background:

• Interferon regulatory factor 4 (IRF4) is a crucial transcription factor in mammalian immune responses.
• Limited understanding of IRF4 function in zebrafish, a key vertebrate model.
• Need for tools to study IRF4a in a living system.

Purpose of the Study:

• To generate transgenic zebrafish with overexpressed irf4a.
• To establish a fluorescently labeled model for tracking IRF4a expression.
• To create a tool for future studies on IRF4a's role in immune responses.

Main Methods:

• Amplification of the zebrafish irf4a coding sequence.
• Construction of a Tol2-based vector for homologous recombination (Tol2-ef1α-irf4a-EGFP).
• Microinjection into zebrafish embryos, screening for fluorescence, and genetic integration confirmation via PCR.

Main Results:

• Successfully generated transgenic zebrafish with systematic overexpression of irf4a.
• Green fluorescence (EGFP) observed in multiple tissues, including spine, eyes, heart, and brain.
• Confirmed stable integration of the irf4a gene into the zebrafish genome across generations.

Conclusions:

• Established a novel transgenic zebrafish line expressing irf4a with ubiquitous green fluorescence.
• This model provides a valuable resource for in vivo studies of IRF4a.
• The transgenic zebrafish will facilitate future research into IRF4a's function in immune responses, particularly concerning inflammation.