Digital Sensing and Molecular Computation by an Enzyme-Free DNA Circuit
Digital Sensing and Molecular Computation by an Enzyme-Free DNA Circuit
复制标题
DOI：
10.1021/acsnano.0c00628
复制
发表时间：
2020-05-26
期刊：
影响因子：
17.1
通讯作者：
Knowles, Tuomas P. J.
中科院分区：
文献类型：
--
作者：
Arter, William E.;Yusim, Yuriy;Knowles, Tuomas P. J.
DNA电路形成了能够进行信号转导和算法计算的可编程分子系统的基础。某些类型的分子程序，如催化发夹组装，使等温，无酶信号放大。然而，在DNA扩增电路中的电流检测极限是适度的，因为灵敏度被非共价体系固有的非催化背景反应导致的信号泄漏抑制。在这里，我们克服了这一挑战，通过优化催化的发夹组件的单分子传感在数字液滴测定。此外，我们证明了在单分子水平上的DNA计算的数字报告，采用ddCHA作为简单的DNA逻辑门的信号转换器。通过在pM浓度下促进分子计算的信号转导，我们的方法可以相对于常规DNA逻辑门将处理密度提高10（4）倍。更广泛地说，我们相信数字分子计算将扩大DNA计算，生物传感和信号放大中的等温放大电路的范围和功效。
DNA circuits form the basis of programmable molecular systems capable of signal transduction and algorithmic computation. Some classes of molecular programs, such as catalyzed hairpin assembly, enable isothermal, enzyme-free signal amplification. However, current detection limits in DNA amplification circuits are modest, as sensitivity is inhibited by signal leakage resulting from noncatalyzed background reactions inherent to the noncovalent system. Here, we overcome this challenge by optimizing a catalyzed hairpin assembly for single-molecule sensing in a digital droplet assay. Furthermore, we demonstrate digital reporting of DNA computation at the single-molecule level by employing ddCHA as a signal transducer for simple DNA logic gates. By facilitating signal transduction of molecular computation at pM concentration, our approach can improve processing density by a factor of 10(4) relative to conventional DNA logic gates. More broadly, we believe that digital molecular computing will broaden the scope and efficacy of isothermal amplification circuits within DNA computing, biosensing, and signal amplification in general.