Honors College Thesis


Improving Bioanalytical Methods of miRNA Analysis and its Applications to Breast Cancer Public Deposited

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  • Micro-ribonucleic acids (miRNA) are becoming an increasingly popular topic of research in hopes of discovering newfound biomarkers and/or therapies for numerous types of cancers and diseases. MiRNA are a type of biomolecule contributing to the progression of multiple cancers and diseases, and such biomolecules are called biomarkers. Current RNA-based biosensors suffer from various flaws such as slow analysis, false signals, sensitivity or the lack of fine-tuned selectivity. The reporter-probe biosensor presented here demonstrates great potential in resolving some of these issues. The biosensor consists of a fluorescent reporter molecule bound to a probe that is more selective for a miRNA than the reporter. The sensor works by simultaneously forming a probe-miRNA target complex and displacing the reporter. The freed reporter is then able to bind to itself using intramolecular forces, forming a hairpin structure inducing a fluorescent signal change. This change in analytical signal can then be measured and related to presence or absence determination and quantification of the miRNA. This innovative miRNA reporter-probe biosensor design is not only capable of reducing false positives, demonstrating selectivity towards three miRNA, but is highly sensitive, a key component to a good biosensor due to the natural low levels (Femtomolar and nanomolar) of miRNA expression in the body. The results of my work have contributed to ongoing studies in the Burrows Lab to one day design an optimal biosensor demonstrating all these feats. Key Words: Biosensor, reporter-probe complex, probe-target complex, hairpin conformation, fluorescence resonance energy transfer (FRET), quenching, false signals, sensitivity, selectivity, and micro-ribonucleic acid (miRNA)
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