Nanopore sensing is developing into a powerful label-free approach to investigate the features of biomolecules at the single-molecule level.When a charged molecule is captured within a nanopore,it modulates the ionic current,which can be recorded in real time to reveal the properties of the target molecule.To date,nanopores have been used to sense a variety of analytes,including DNA,RNA,proteins,enzymes,small molecules,cancer cells,and metal ions,and can also provide information on biomolecular structures.In this review,we highlight the progress made in nanopore sensing over the past five years (2016-2020),and provide an outlook on future developments and directions in the field.