| An innovative therapeutic nano device---intelligent nanomolecular (NM) device is presented in this dissertation. The NM device consists of two modules: the diagnosis module is a hairpin oligonucleotide that can recognize the specific mRNA biomarker and the therapy module consists of a drug and an inhibitor of the drug. Upon recognition of the disease-indicating biomarker, the diagnosis module will change its conformation, thus leading to activation or deactivation of the drug in the diagnosis module. The feasibility of the INM design concept was proved in a model system with both thermodynamics-based predications and experiments. In the model system, GFP mRNA is the biomarker and thrombin is the disease-causing molecule. Four INM devices, whose therapy module consists of an anti-thrombin aptamer and a complementary sequence to the aptamer, were designed. The comparison between these different designs revealed other important design considerations in addition to the thermodynamics. The possibility to diversify and rationalize the design of the INM device was indirectly demonstrated with an aptamer-based optical sensor that was derived from one of extended INM design concepts. Furthermore, a few critical issues involved in the intracellular testing of the INM device, including intracellular delivery, stability, and specificity, were also investigated with molecular beacons in a micro fluidic cell culture system. |
