Recent developments in the characterization of nucleic acid hybridization kinetics
Recent developments in the characterization of nucleic acid hybridization kinetics
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DOI：
10.1016/j.cobme.2021.100305
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发表时间：
2021-07-12
影响因子：
3.9
通讯作者：
Yeh, Hsin-Chih
中科院分区：
文献类型：
--
作者：
Chen, Yuan-, I;Sripati, Manasa P.;Yeh, Hsin-Chih
核酸杂交(NAS)是驱动许多细胞过程的基本分子机制，使新的生物技术和治疗成为可能。然而，现有的测量NAS杂交动力学的方法要么是在系综水平上进行的，要么是限于非天然生理条件下进行的。三维单分子跟踪技术的最新进展打破了这些限制，允许在活的哺乳动物细胞内自由扩散的单个寡核苷酸上观察到多个退火和融化事件。本文综述了在单分子水平和活细胞中测量NA杂交动力学的各种方法，并简要介绍了活细胞杂交动力学测量中尚未解决的挑战和机遇。还描述了NA动力学测量和生物技术所取得的重要发现，这些发现可以随着对杂交动力学的深入了解而得到改进。
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 threedimensional 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.