Single-Molecule Methods for Characterizing Different DNA Higher-Order Structures

DNA is considered to be not only a carrier of the genetic information of life but also a highly programmable and self-assembled nanomaterial. Different DNA structures are related to their biological and chemical functions. Hence, understanding the physical and chemical properties of various DNA structures is of great importance in biology and nanochemistry. However, the bulk assay ignores the heterogeneity of DNA structures in solution. Single-molecule methods are powerful tools for observing the behavior of individual molecules and probing the high heterogeneity of free energy states. In this review, we introduce single-molecule methods, including single-molecule detection and manipulation methods, and discuss how these methods can be conducive to measuring the molecular properties of single-/ double-stranded DNA (ss/dsDNA), DNA higher-order structures, and DNA nanostructures. We conclude by providing a new perspective on the combination of DNA nanotechnology and single-molecule methods to understand the biophysical properties of DNA and other bio-matter and soft matter.(c) 2022 THE AUTHORS. Published by Elsevier LTD on behalf of Chinese Academy of Engineering and Higher Education Press Limited Company. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Automated summary

DNA is not only a carrier of genetic information, but also a programmable and self-assembled nanomaterial. Different DNA structures are closely related to their biological and chemical functions. Single-molecule methods provide a powerful tool for studying the behavior and heterogeneity of individual DNA molecules. This review discusses the application of single-molecule methods in measuring the properties of DNA structures, including single-stranded/double-stranded DNA, DNA higher-order structures, and DNA nanostructures. The combination of DNA nanotechnology and single-molecule methods offers a new perspective for understanding the biophysical properties of DNA and other bio-matter and soft matter.