Recent developments in the characterization of nucleic acid hybridization kinetics. (September 2021)

• Record Type: Journal Article
• Authors: Chen, Yuan-I; Sripati, Manasa P.; Nguyen, Trung Duc; Chang, Yin-Jui; Kuo, Yu-An; Hong, Soonwoo; Yeh, Hsin-Chih
• Is Part Of: Current opinion in biomedical engineering. Volume 19(2021)
• Publication Date: 2021-09
• Subjects: Nucleic acids -- Hybridization kinetics -- Single-molecule detection -- Single-molecule tracking -- Live-cell studies; Biomedical engineering -- Periodicals; 610.28
• DOI: 10.1016/j.cobme.2021.100305
• Languages: English
• ISSNs: 2468-4511

Abstract

Abstract: Hybridization of nucleic acids (NAs) is a fundamental molecular mechanism that drives many cellular processes and enables new biotechnologies as well as therapeutics. However, existing methods that measure hybridization kinetics of NAs are either performed at the ensemble level or constrained to non-native physiological conditions. Recent advances in three-dimensional single-molecule tracking techniques break these limitations by allowing multiple annealing and melting events to be observed on a single oligonucleotide freely diffusing inside a live mammalian cell. This review provides an overview of diverse approaches to measuring NA hybridization kinetics at the single-molecule level and in live cells, and concludes with a synopsis of unresolved challenges and opportunities in the live-cell hybridization kinetics measurements. Important discoveries made by NA kinetics measurements and biotechnologies that can be improved with a deeper understanding of hybridization kinetics are also described.