RNA-lipid · Characterisation and design of RNA-lipid interactions for RNA therapeutics

RNA is a ubiquitous biomolecule with a broad range of activities crucial for keeping cells alive. Understanding how RNA bioactivity is controlled is vital for elucidating its roles and applications in molecular biology. RNA, in order to maintain a correct activity patterns, has to be organized. Despite sufficient knowledge about RNA spatial organization within cells via RNA-protein interaction-based processes not much is known about other mechanisms. Revealing the RNA-localization patterns without RNA-binding proteins would create the new branch of interactomics which might have significant implications in the molecular biology and RNA-based therapeutics development.Recently I described that lipid membranes can act as a scaffold for RNAs in a sequence and structure dependent binding process leading to the changes of the RNA activity in vitro. Here, I propose that lipid membranes act as a selective RNA scaffold in vivo. In the following project I will develop a library of membrane-binding RNAs using in vitro selection approach. I will characterize the mechanisms, selectivity, and stability of RNA-lipid binding in bulk and in nano-scale level using self-developed binding assays and super-resolution microscopy, respectively. Furthermore, obtained lipid-binding RNAs are going to be artificially introduced to the existing mRNAs in order to spatially organize protein expression both in vitro and in vivo. The RNA RNA-lipid encapsulation efficiency will be characterized using recently developed single-particle profiling platform. Lastly, I will use gathered knowledge to livetrack the uptake and improve the RNA-based therapeutics (i.e. RNA-lipid nanoparticles) in order to increase the RNA stability and decrease the toxicity of the formulation.