The iGEM Laboratory of OUC-China, College of Marine Life Sciences, Ocean University of China, Qingdao, China.
Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, Shandong, China.
Bioengineered. 2022 Jan;13(1):1235-1248. doi: 10.1080/21655979.2021.2020493.
To date, several different types of synthetic genetic switches, including riboregulators, riboswitches, and toehold switches, have been developed to construct AND, OR, NOT, NAND, NOR, and NOT IMPLICATION (NIMP) gates. The logic gate can integrate multiple input signals following a set of algorithms and generate a response only if strictly defined conditions are met. However, there are still some logic gates that have not been implemented but are necessary to build complex genetic circuits. Here, based on the toehold switches and three-way-junction (3WJ) repressors, we designed two novel biological Boolean logic gates of IMPLICATION (IMP) and XOR. Subsequently, the outputs of these two logic gates were characterized by fluorescence analysis, indicating that they can achieve the truth tables of logical gates. Furthermore, the fluorescence intensity under the logical TRUE condition was significantly higher than under the logical FALSE condition, suggesting the high dynamic range of the ON/OFF ratios. Because of the programmability of synthetic RNA switches, the constructed RNA logic gates could serve as elementary units to build a versatile and powerful platform for translational regulation and RNA-based biological computation.
迄今为止，已经开发出了几种不同类型的合成遗传开关，包括核糖开关、核糖调控器和链置换开关，用于构建与门、或门、非门、与非门、或非门和蕴涵门（NIMP）。逻辑门可以根据一组算法集成多个输入信号，并且仅在严格定义的条件得到满足时才会产生响应。然而，仍然有一些逻辑门尚未实现，但对于构建复杂的遗传电路是必要的。在这里，我们基于链置换开关和三链结（3WJ）抑制剂，设计了两种新型的生物布尔逻辑门，蕴涵门（IMP）和异或门（XOR）。随后，通过荧光分析对这两个逻辑门的输出进行了表征，表明它们可以实现逻辑门的真值表。此外，逻辑 TRUE 条件下的荧光强度明显高于逻辑 FALSE 条件下的荧光强度，这表明其开启/关闭比值的动态范围较高。由于合成 RNA 开关的可编程性，所构建的 RNA 逻辑门可以作为基本单元，为基于翻译调控和 RNA 的生物计算构建一个通用且强大的平台。
