Date of Graduation
Master of Science in Cell and Molecular Biology
splice switching oligonucleotides, SSO, BRAF, delivery, cancer, nanotechnology, biomedical, transfection, drug delivery, bioinformatics
Medical Molecular Biology
Splice switching oligonucleotides (SSOs) are an emerging antisense RNA drug class with the ability to therapeutically modify gene expression. A wide variety of chemical modifications have been devised to try to increase the activity and stability of SSOs. Also, as with most nucleic acid therapeutics, delivery into the cell is the major hurdle for in vivo and clinical applications. As a result, an array of nano delivery vehicles are being developed to overcome this obstacle. However, it is difficult to find a practical assay to measure splice switching activity. Here, I characterize a model delivery system that can be used as a convenient, high-throughput assay to quantitatively measure the functional delivery and splicing redirection in a human melanoma cell line. Utilizing this system, I screen a variety of nanomaterials and discover that cobalt oxide is capable of inherent, unassisted SSO delivery into cancer cells. I also design a set of novel SSOs using a genomics based approach to target BRAF, a prevalent oncogene in human cancer, and assess their therapeutic potential against human melanoma.
© John Michael Dean
Dean, John Michael, "Splice Switching Oligonucleotides: Model System Characterization, Nanomaterial Delivery, and Rational Design for Targeting Melanoma at the RNAa Level" (2014). MSU Graduate Theses. 1860.