Label-free assembly of RNA-containing viruses, one particle at a time

Self-assembly of viral particles remains a poorly understood process. What typically is known is the starting state, i.e. what the individual proteins and the genome look like, as well as the end state, i.e. the structure of a complete virus. However, what happens in between is largely unclear. The reason for that is that it is difficult to capture the intermediate assembly states with standard techniques. In this project, this problem will be tackled by using state-of-the-art single molecule techniques that are able to follow the early steps in assembly of individual particles. This means that viral assembly initiation can be observed particle by particle. While traditional ensemble averaging techniques are very useful to decipher the overall thermodynamics of viral assembly, they do not account for individual variations. Single particle techniques have an exactly opposite approach as these variations are captured, because each individual particle is followed. This will allow for understanding the variability in assembly and lay bare possible pathways of this process. In order to interfere as minimally as possible with assembly, label-free techniques are used that do not influence or alter the particle-particle interactions. In this manner, assembly initiation of RNA containing viruses will be studied. The new insights into assembly that will be generated are essential for constructing fundamental knowledge on this poorly understood process. The results will additionally be available for using viral nanoparticles in nanotechnology and medicine, as well as for fighting viruses more efficiently.