Key Laboratory of Green Chemistry & Technology of Ministry of Education, College of Chemistry, Sichuan University, Chengdu, Sichuan 610064, China.
Department of Chemistry, Brock University, 1812 Sir Isaac Brock Way, St. Catharines, Ontario L2S 3A1, Canada.
J Am Chem Soc. 2024 Mar 13;146(10):6516-6521. doi: 10.1021/jacs.3c09242. Epub 2024 Feb 27.
Regulatory modules for controlling the kinetics of toehold-mediated strand displacement (TMSD) play critical roles in designing dynamic and dissipative DNA chemical reaction networks (CRNs) but are hardwired into sequence designs. Herein, we introduce antitoehold (At), a plug-and-play module for reversible and continuous tuning of TMSD kinetics by temporarily occupying the toehold domain via a metastable duplex and base stacking. We demonstrate that kinetic control can be readily activated or deactivated in real time for any TMSD by simply adding At or anti-At. Continuous tuning of TMSD kinetics can also be achieved by altering the concentration of At. Moreover, the simple addition of At could readily reprogram existing TMSDs into a pulse-generation DNA CRN with continuous tunability. Our At approach also offers a new way for engineering continuously tunable DNA hybridization probes, which may find practical uses for discriminating clinically important mutations. Because of the simplicity, we anticipate that At will find wide applications for engineering DNA CRNs with diverse dynamic and dissipative behaviors, and DNA hybridization probes with tunable affinity and selectivity.
调控模块用于控制引发链置换（TMSD）的动力学在设计动态和耗散 DNA 化学反应网络（CRN）方面起着至关重要的作用，但这些调控模块是通过序列设计硬连线实现的。在此，我们引入了反引发（At），这是一个即插即用的模块，通过亚稳态双链体和碱基堆积暂时占据引发序列域，从而实现 TMSD 动力学的可逆和连续调节。我们证明，通过简单地添加 At 或 anti-At，可以实时轻松地激活或停用任何 TMSD 的动力学控制。通过改变 At 的浓度，也可以实现 TMSD 动力学的连续调节。此外，只需添加 At，就可以将现有的 TMSD 轻松重新编程为具有连续可调性的脉冲生成 DNA CRN。我们的 At 方法还为工程连续可调谐 DNA 杂交探针提供了一种新方法，这可能在区分临床上重要的突变方面具有实际用途。由于其简单性，我们预计 At 将广泛应用于具有不同动态和耗散行为的 DNA CRN 工程以及具有可调谐亲和力和选择性的 DNA 杂交探针工程。
