RNA Polymerase Transcription Elongation Pasuing and Termination Investigated at Super Resolution with the Mspa Nanopore

We have applied a new single molecule technique named SPRNT (Single-molecule Picometer Resolution Nanopore Tweezers) to analyze the translocation of single E. coli RNA polymerase (RNAP) complexes during transcription. Inspired by nanopore DNA sequencing, SPRNT enables observation of motor protein movement along nucleic acids with sub-nucleotide spatial resolution, sub-millisecond temporal resolution, and the ability to apply assisting and opposing forces. In addition, the sequence in the RNAP complex is provided by nanopore sequencing, allowing for study of sequence-dependent translocation behaviors.