Item type:Thesis, Open Access

Optochemical Regulation of mRNA-Based Processes

Loading...
Thumbnail Image

relationships.isAuthorOf

Publisher

Philipps-Universität Marburg

item.page.supervisor-of-thesis

Abstract

Optochemical tools serve as potent instruments for the modulation of RNA-based processes. This thesis introduces innovative development in the optochemical control of RNA splicing and translation regulation, utilizing photoswitchable small molecules and peptide nucleic acids (PNAs). In one part of this thesis (Chapter 3.1 and 3.3), an approach involving visible light-responsive peptide nucleic acids (PNAs) for the manipulation of oligonucleotide hybridization was introduced. The synthesis of these PNAs involved the conjugation with photoswitches (ortho-tetrafluoroazobenzene (oF4Azo), hemithioindigo, and spiropyran). The subsequent synthesis and detailed photochemical characterization were expounded upon. The studies revealed the promising attributes of oF4Azo-PNA conjugates, including rapid and reversible isomerization, remarkable thermal stability, high isomer conversion, and sensitivity to visible-light irradiation. Through melting experiments and strand-displacement assays, the investigation explored light-controlled oligonucleotide hybridization, showing modest yet discernible differences induced by light. Combining oF4Azo-PNAs with cell-penetrating peptides (CPPs) yielded light-responsive antisense agents for use in living cells. With distinct goals of redirecting mRNA splicing and mimicking microRNA to inhibit cancer cell growth, the PNA-CPPs demonstrated favorable outcomes. However, it was observed that the degree of light controllability was limited. The subsequent development of cyclic PNAs was introduced, alongside various synthetic approaches. Studies indicated that the ring size influenced oligonucleotide hybridization. Another part of this thesis (Chapter 3.2) delved into the development of reversible mRNA splicing regulation through photoswitchable small molecules. The synthesis and comprehensive photochemical analysis of synthesized molecules were meticulously detailed. The best compound, Me-oF2Azo-BOX, exhibited rapid and reversible photoisomerization, good thermal stability, and significant differences in enhancing exon inclusion in splicing between photostationary states. This studies underscored its capability to achieve real-time conditional control at both mRNA and protein levels, which could be reversibly regulated by light. Molecular mechanisms of action were further elucidated.

Review

Metadata

show more
Zhang, Lei: Optochemical Regulation of mRNA-Based Processes. : Philipps-Universität Marburg 2024-07-22. DOI: https://doi.org/10.17192/z2023.0679.