A novel strategy for selection of allosteric ribozymes yields RiboReporter sensors for caffeine and aspartame |
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Authors: | Ferguson Alicia Boomer Ryan M Kurz Markus Keene Sara C Diener John L Keefe Anthony D Wilson Charles Cload Sharon T |
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Institution: | Archemix Corporation, 1 Hampshire Street, Cambridge, MA 02139, USA. |
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Abstract: | 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™ 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. |
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