Single-Molecule Picometer Resolution Nanopore Tweezers Resolution

Single-molecule Picometer Resolution Nanopore Tweezers (SPRNT) is a new technique for observing the motion of single motor enzymes on nucleic acid tracks with more than an order of magnitude improvement in spatiotemporal resolution over existing single-molecule technologies such as optical tweezers. Because SPRNT is derived from nanopore DNA sequencing, it also provides a readout of the DNA sequence moving through the enzyme, allowing us to query how nucleic acid sequence regulates the motion of motor enzymes. We discuss how SPRNT works in the context of helicase and RNA polymerase systems to which we have applied SPRNT, and present results including a full kinetic model relating ATP-hydrolysis steps to sub-nucleotide helicase motion and observation of RNA polymerase pausing and termination at unprecedented resolution.