Experimental Validation of Signal Dependent Operation in Whiplash PCR

摘要

Whiplash PCR (WPCR), which implements self-directed operation, programmed within a single DNA molecule, is a potential candidate for both mathematical and biological applications. However, WPCR-based methods are known to suffer from a serious efficiency problem called back-hybridization (BH). Previously, we proposed and partially validated a new rule-protect operation to abolish BH. In this work, we experimentally demonstrate the ability of rule-protect to drive multi-step WPCR. Successful implementation of isothermal operation at physiological temperatures is an essential benchmark for biological applications. We also propose the use of rule-protect for external signalling to control computational operation. Consequently, signal-dependent self-directed operation, which is conceptually new to DNA computing, is achieved. The present architecture, provided with sensing ability, allows a composite system design layering computational reactions, and would be suitable for functioning as the central processing unit of this system.