Investigating lipid formulations for efficient RNA delivery using zebrafish models

Abstract

Lipid formulations have demonstrated strong potential for nucleic acid delivery,

particularly with the success of COVID-19 vaccines. Despite these advances, many

aspects of lipid formulations remain unclear. For instance, how these particles enter

cells or how their composition might be tuned for targeted delivery to specific tissues or cell types still needs to be fully understood. This thesis aims to explore the underlying mechanisms that influence the in vivo behavior of lipid nanoparticles, using zebrafish embryos as a model. Due to their transparency, zebrafish embryos offer a unique opportunity to observe the dynamic interactions between fluorescentlabeled nanoparticles and living cells in real time. They also enable the quantification of mRNA translation into fluorescent proteins, as well as the effects of gene silencing. Furthermore, by employing CRISPR/Cas9 to generate knockout zebrafish lines, we can evaluate receptor-specific interactions in a living organism. The overarching aim of this work is to gain mechanistic insight that can inform the design of more effective and targeted lipid formulations.