SpeeDx Pty Ltd, Eveleigh, New South Wales, Australia 2015.
J Am Chem Soc. 2010 Jan 27;132(3):1051-9. doi: 10.1021/ja9076777.
To increase the versatility and utility of nucleic acid enzymes, we developed multicomponent complexes, known as MNAzymes, which produce amplified "output" signals in response to specific "input" signals. Multiple oligonucleotide partzymes assemble into active MNAzymes only in the presence of an input assembly facilitator such as a target nucleic acid. Once formed, MNAzymes catalytically modify a generic substrate, generating an amplified output signal that heralds the presence of the target while leaving the target intact. We demonstrated several applications including sensitive, isothermal target detection; discrimination of polymorphisms; and highly specific monitoring of real-time polymerase chain reaction (PCR). Furthermore, we showed their capacity to function as molecular switches and to work in series to create a molecular cascade. The modular nature of MNAzymes, together with the separation of input and output functionalities, provides potential for their integration into diverse devices such as diagnostic biosensors, molecular computers, and/or nanoscale machines.
为了提高核酸酶的多功能性和实用性，我们开发了多组分复合物，称为 MNAzymes，它们可以对特定的“输入”信号产生放大的“输出”信号。只有在存在输入组装促进剂（如靶核酸）的情况下，多个寡核苷酸部分酶才能组装成活性 MNAzymes。形成后，MNAzymes 催化修饰通用底物，产生放大的输出信号，预示着靶标存在，同时保持靶标完整。我们展示了几种应用，包括敏感、等温的靶标检测；多态性的区分；以及实时聚合酶链反应（PCR）的高度特异性监测。此外，我们还表明它们可以作为分子开关发挥作用，并串联工作以创建分子级联。MNAzymes 的模块化性质以及输入和输出功能的分离，为它们在诊断生物传感器、分子计算机和/或纳米级机器等各种设备中的集成提供了潜力。
