Main description:

This book describes the development of novel protein-RNA-binding assays and their applications in a high-throughput manner for the identification of small-molecule modulators of protein-RNA interactions to treat cancer and COVID-19.

Modulating protein-RNA interactions with small molecules is expected to provide novel biological insights of the interrelation of diseases with the protein-RNA interactome. The modulations may also be exploited therapeutically. For these reasons, the development of a simple, reliable, and sensitive protein-RNA-binding assay is necessary for high-throughput screening to discover new effective chemical entities capable of acting on diverse protein-RNA interactions. This book discusses the discovery of small-molecule modulators targeting protein-RNA interactions that are potentially valuable to treat cancer and COVID-19 by constructing novel high-throughput screening methods. The results of this dissertation provide valuable insights into the regulation of protein-RNA interactions in chemical biology and drug development.

Contents:

Chapter 1. Introduction............................................................................ 1

1.1. Protein-RNA interaction.............................................................. 1

1.2. Therapeutic potentials of modulating PRI.................................... 1

1.3. Chemical biology approaches for the discovery of PRI modulator

............................................................................................................. 3

1.4. Aims and scope of the thesis........................................................ 5

1.5. References.................................................................................... 5

Chapter 2. Identification of Small-Molecule Inhibitors of Oncogenic Lin28-Let-7 Interaction 8

2.1. Discovery of a small-molecule inhibitor of protein-microRNA interaction using binding assay with a site-specifically labeled Lin28..................................................................... 8

2.1.1. Introduction..................................................................... 8

2.1.2. Results and discussion................................................... 11

2.1.3. Conclusion..................................................................... 26

2.1.4. Experimental section..................................................... 27

2.1.5. References..................................................................... 37

2.2. Restoring let-7 microRNA biogenesis using a small-molecule inhibitor of the protein-RNA interaction................................................................................................................... 41

2.2.1. Introduction................................................................... 41

2.2.2. Results and discussion................................................... 43

2.2.3. Conclusion..................................................................... 53

2.2.4. Experimental section..................................................... 53

2.2.5. References..................................................................... 58

Chapter 3. Discovery of Small-Molecule Modulators of Protein-RNA Interactions by Fluorescence Intensity-Based Binding Assay............................................................... 61

3.1. Introduction................................................................................ 61

3.2. Results and discussion................................................................ 63

3.3. Conclusion.................................................................................. 76

3.4. Experimental section.................................................................. 76

3.5. References.................................................................................. 85

Chapter 4. Identification of Protein-RNA Interaction Modulators for Treating COVID-19 88

4.1. Harnessing stress granule formation by small molecules to inhibit the cellular replication of SARS-CoV-2......................................................................................................... 88

4.1.1. Introduction................................................................... 88

4.1.2. Results and discussion................................................... 89

4.1.3. Conclusion................................................................... 103

4.1.4. Experimental section................................................... 104

4.1.5. References.................................................................... 110

4.2. Discovery of a small-molecule degrader of SARS-CoV-2 Nsp1 by autophagic pathway 112

4.2.1. Introduction.................................................................. 112

4.2.2. Results and discussion................................................. 113

4.2.3. Conclusion................................................................... 119

4.2.4. Experimental section................................................... 119

4.2.5. References 121