Ferguson Alicia, Boomer Ryan M, Kurz Markus, Keene Sara C, Diener John L, Keefe Anthony D, Wilson Charles, Cload Sharon T
Archemix Corporation, 1 Hampshire Street, Cambridge, MA 02139, USA.
Nucleic Acids Res. 2004 Mar 16;32(5):1756-66. doi: 10.1093/nar/gkh336. Print 2004.
We have utilized in vitro selection technology to develop allosteric ribozyme sensors that are specific for the small molecule analytes caffeine or aspartame. Caffeine- or aspartame-responsive ribozymes were converted into fluorescence-based RiboReporter trade mark sensor systems that were able to detect caffeine or aspartame in solution over a concentration range from 0.5 to 5 mM. With read-times as short as 5 min, these caffeine- or aspartame-dependent ribozymes function as highly specific and facile molecular sensors. Interestingly, successful isolation of allosteric ribozymes for the analytes described here was enabled by a novel selection strategy that incorporated elements of both modular design and activity-based selection methods typically used for generation of catalytic nucleic acids.
我们利用体外筛选技术开发了对小分子分析物咖啡因或阿斯巴甜具有特异性的变构核酶传感器。对咖啡因或阿斯巴甜有反应的核酶被转化为基于荧光的RiboReporter商标传感器系统，该系统能够在0.5至5 mM的浓度范围内检测溶液中的咖啡因或阿斯巴甜。这些依赖于咖啡因或阿斯巴甜的核酶读取时间短至5分钟，可作为高度特异性且便捷的分子传感器。有趣的是，此处所述分析物的变构核酶的成功分离是通过一种新颖的筛选策略实现的，该策略结合了模块化设计和通常用于生成催化核酸的基于活性的筛选方法的要素。
