Institut für Biochemie, Ernst-Moritz-Arndt-Universität Greifswald, Felix-Hausdorff Str. 4, 17487, Greifswald, Germany.
Adv Biochem Eng Biotechnol. 2020;170:21-35. doi: 10.1007/10_2017_21.
Engineering of nucleic acids has been a goal in research for many years. Since the discovery of catalytic nucleic acids (ribozymes and DNAzymes), this field has attracted even more attention. One reason for the increased interest is that a large number of ribozymes have been engineered that catalyze a broad range of reactions of relevance to the origin of life. Another reason is that the structures of ribozymes or DNAzymes have been modulated such that activity is dependent on allosteric regulation by an external cofactor. Such constructs have great potential for application as biosensors in medicinal or environmental diagnostics, and as molecular tools for control of cellular processes. In addition to the development of nucleic acid enzymes by in vitro selection, rational design is a powerful strategy for the engineering of ribozymes or DNAzymes with tailored features. The structures and mechanisms of a large number of nucleic acid catalysts are now well understood. Therefore, specific design of their functional properties by structural modulation is a good option for the development of custom-made molecular tools. For rational design, several parameters have to be considered, and a number of tools are available to help/guide sequence design. Here, we discuss sequence, structural and functional design using the example of hairpin ribozyme variants to highlight the challenges and opportunities of rational nucleic enzyme engineering.
多年来，核酸的工程设计一直是研究领域的目标。自发现催化性核酸（核酶和脱氧核酶）以来，该领域吸引了更多的关注。兴趣增加的原因之一是已经设计出了大量的核酶，它们可以催化与生命起源相关的广泛的反应。另一个原因是核酶或脱氧核酶的结构已经被调节，使得活性依赖于外部辅助因子的变构调节。这些构建物在医学或环境诊断中的生物传感器应用以及作为控制细胞过程的分子工具方面具有巨大的潜力。除了通过体外选择开发核酸酶外，理性设计也是具有定制特征的核酶或脱氧核酶工程的有力策略。大量核酸催化剂的结构和机制现在已经得到很好的理解。因此，通过结构调节对其功能特性进行特定设计是开发定制分子工具的一个很好的选择。对于合理的设计，需要考虑几个参数，并且有许多工具可用于帮助/指导序列设计。在这里，我们将以发夹核酶变体为例讨论序列、结构和功能设计，以突出理性核酸酶工程设计的挑战和机遇。
