Inspired by the biological processes of molecular recognition and transportation across membranes, nanopore techniques have evolved in recent decades as ultrasensitive analytical tools for individual molecules. In particular, nanopore-based single-molecule DNA/RNA sequencing has advanced genomic and transcriptomic research due to the portability, lower costs and long reads of these methods. Nanopore applications, however, extend far beyond nucleic acid sequencing. In this Review, we present an overview of the broad applications of nanopores in molecular sensing and sequencing, chemical catalysis and biophysical characterization. We highlight the prospects of applying nanopores for single-protein analysis and sequencing, single-molecule covalent chemistry, clinical sensing applications for single-molecule liquid biopsy, and the use of synthetic biomimetic nanopores as experimental models for natural systems. We suggest that nanopore technologies will continue to be explored to address a number of scientific challenges as control over pore design improves.

In recent years, single molecule technology has experienced a rapid growth, with exciting developments in fundamental research and real-world applications. Detecting and studying biological phenomena on the
single molecule level requires a unique synergy between researchers working on instrumentation, physics, and the life sciences. In the iScience special issue ‘‘Single Molecule Technology – From Biotechnology to
Biomedical Applications’’, guest edited by Amit Meller and Chirlmin Joo (Figure 1), we are highlighting a variety of research on nanopore technology, single molecule fluorescence, and a selection of other ultra-
sensitive detection methods. More content in the special issue can be found here: https://www.sciencedirect.com/journal/iscience/special-issue/10PGSBV55N0. The guest editors in this backstory share their thoughts on what is currently exciting in the field, and the advances they think will make an impact in the near future.