Stall force measurement of the kinesin-3 motor KIF1A using a programmable DNA origami nanospring

DNA origami technology is a method for designing and constructing nanoscale structures using DNA, and it is being applied across various fields. This technology was advanced by developing the nanospring (NS), a fluorescently visible molecular spring that quantifies forces through its extension and has been used to measure myosin-generated forces. This study aims to measure the force exerted by the kinesin-3 motor protein KIF1A, mutations of which cause KIF1A-associated neurological disorder (KAND) and are associated with reduced force and motility. Unlike kinesin-1, KIF1A detaches easily under perpendicular loads, which can occur in optical tweezers experiments. By applying force parallel to the microtubule using the NS, we were able to precisely measure the stall force even for KAND mutants, for which such measurements are typically challenging. This result highlights the potential of the NS as a new tool for force spectroscopy in biophysics.