Using zebrafish as a model to investigate nanoparticle-based carriers for drug delivery and gene editing

Nanoparticles (NPs) are used across a wide range of fields, including medicine, electronics, energy, and environmental science. In medicine, they are employed for drug delivery, imaging. Zebrafish (Danio rerio) has been extensively used as a model organism to investigate NPs biological effects and potential applications in both biomedical research and environmental science. The aim of this work, in the frame of the PNRR project (National Center for Gene Therapy and Drugs based on RNA Technology; Identification of models to assess delivery efficiency in zebrafish embryos), is to expand knowledge and enhance the potential of NPs for drug delivery through their tropism, particularly toward the heart and endothelial cells of blood vessels. Moreover, we are paving the way to generate a zebrafish transgenic line capable of ubiquitously expressing the Cas9 protein. This approach will allow for testing whether the guide RNAs (gRNAs) designed to knock out target genes are delivered to specific tissues based on different types of NPs carriers used. To monitor NPs uptake and migration, 48 hours post-fertilization zebrafish larvae have been injected through different routes. Daily observation to assess larval viability and any toxic effects has been performed, while the position of the NPs have been tracked using fluorescence and confocal microscopy. Encouraging results about the safety and tropism of the tested NPs have been recorded. However, further investigation into the composition and the formulation of these NPs is needed to improve their bioavailability and to evaluate if they can be used for medical purposes