One of the great scientific questions of molecular biology is how RNA and protein molecules fold into their final functional shapes. Even though some progress has been made, we are still very far from being able to predict the folding of complex molecular machines such as the ribosome, and even farther from being able to rationally design similar structures. In the COFOLD project we will study how RNA molecules fold during synthesis (co-transcriptional folding) using rational design of RNA folding pathways (RNA origami) as a key tool to gain detailed insight into this process. Cryo-electron microscopy will be used to characterize the 3-dimensional structure of co-transcriptional folding products and single-molecule fluorescence microscopy will be used to determine core kinetic parameters of folding events. Folding in cells will be investigated by cryo-electron tomography and fluorescence microscopy. The resulting insights will be used to improve RNA design algorithms and computer-aided design software to reliably predict and design co-transcriptional folding. Improved predictions of RNA folding in space and time will lead to a better understanding of the complex RNA assemblies found in biology and to an extraordinary ability to design functional RNAs, which will be of great importance for both medicine and synthetic biology.
