Comprehensive insights into molecular techniques and applications for genetic modification and targeted mutagenesis in zebrafish

The zebrafish ( Danio rerio ) (ZF) is an emerging model organism in biomedical research. A high degree of evolutionary conservation exists between zebrafish and humans. This helps researchers understand the molecular mechanisms underlying vertebrate development and human diseases. As a genetic model organism, ZF has several benefits, including a fully sequenced genome, rapid external embryonic development with high transparency, a huge number of year-round progenies, and molecular accessibility for genetic modification. The current ZF genome sequence data, combined with high-throughput sequencing methods, facilitate the identification of specific mutations. One of its most notable features is the ZF model’s ability to provide real-time, live imaging of numerous biological processes, including bone development and repair. ZF genetic engineering has become a powerful tool for researching gene function, modeling disease, and drug discovery. This review highlights the shift from gene disruption to precise and reversible control of gene activity by combining traditional mutagenesis techniques with cutting-edge genome-editing tools and newly developed CRISPR-based regulatory systems [CRISPR interference and activation (CRISPRi/a)]. Recent developments such as multiplex genome editing and F 0 CRISPR screening are also covered, along with their uses in aquaculture, disease modeling, and functional genomics. This review will examine novel methods of ZF genetic modification, emphasizing current developments in the field.