Comparative analysis of novel nanoparticle-based gene delivery with conventional methods in fish and crustacean embryos

Efficient gene delivery is critical for gene editing applications in aquatic animals, yet achieving effective transfection remains challenging due to embryo-specific structural barriers. This study systematically compared four gene delivery methods—microinjection, electroporation, lentiviral vectors, and TAT-modified PEG- b -PLGA (TNP) nanoparticles—in embryos of two representative aquatic species: zebrafish ( Danio rerio ) and shrimp ( Penaeus monodon ). Microinjection and electroporation showed relatively high transfection efficiencies in D. rerio but resulted in elevated embryo mortality. Conversely, these methods were ineffective and caused severe mortality in P. monodon embryos. Lentiviral vectors failed entirely in both species, causing 100% embryo lethality. Notably, TNP nanoparticles significantly improved transfection efficiency while reducing mortality rates in both D. rerio and P. monodon embryos. The findings highlight TNP nanoparticles as a superior, promising gene delivery platform suitable for aquatic animal research, although further optimization is required to enhance efficacy and minimize lethality across species.