Monari, L.; Braun, I.; Poppleton, E.; Göpfrich, K.

To realize the full potential of RNA nanotechnology and RNA origami, user-friendly design tools are needed. Here, we present pyFuRNAce, an open-source, Python-based software package with a graphical user interface that enables the design of complex RNA nanostructures. PyFuRNAce integrates the entire RNA origami workflow - from motif definition and blueprint design to sequence generation and primer selection - into a single, user-friendly platform. Built around a motif-based assembly paradigm, the software enables users to create and modify custom nanostructures through an intuitive web interface with streamlined design steps and real-time 3D visualization. By consolidating multiple design stages into a unified environment, pyFuRNAce reduces the entry barrier for RNA nanotechnology and accelerates the development of functional RNA origami structures for applications in medicine, biotechnology, and synthetic biology.