Browsing by Subject "Quinine"

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    Quinine Copolymer Reporters For Enhanced Gene Editing And Raman Imaging
    (2022-01) Van Bruggen, Craig
    After decades of development, gene therapy has finally reached the forefront of medicine and has led to new cures for genetic disorders and the development of life-saving vaccines. The field has been buoyed by the development of more precise and user-friendly targeted nucleases, such as those used for clustered regularly interspersed palindromic repeats (CRISPR)-based editing. These useful gene-editing technologies, however, are still stymied by the challenge of delivering exogenous nucleic acids and proteins into the cells of interest. The emerging gene therapy industry is investing heavily in developing more efficient and safe non-viral vehicles as alternatives to costly and immunogenic viral vectors. Cationic polymers are promising non-viral vectors due to their manufacturing scalability, their chemical stability, and their synthetic tunability. Improvements in delivery efficiency are necessary, however, for widespread adoption of polymeric vehicles for gene therapy. One challenge in improving performance, however, is the difficulty and limited methodology for elucidating the intracellular mechanics of polymeric vehicles. In this thesis, I describe my research focused on the development of a novel quinine-containing polymer, called a Quinine Copolymer Reporter (QCR), that enhanced transient transfections of cultured cells with plasmids and improved gene editing of cultured cells through the simultaneous delivery of the CRISPR-associated protein Cas9 and DNA donor template. In addition, I describe collaborative research performed with colleagues in the research group of Prof. Renee Frontiera that characterized a band in quinine’s Raman spectrum that is diagnostic of its chemical environment. Using this chemical sensitivity in conjunction with Raman microscopic imaging, we help elucidated the intracellular unpackaging mechanisms of the QCR-nucleic acid complexes.