This volume provides a concise and technical discussion of recently developed approaches to overcome challenges, such as pharmacodynamics and pharmacokinetics difficulties, in miRNA drug discovery. These strategies cover anti-sense agents targeting miRNA that are applied in advanced formulations or are chemically optimized to increase delivery; small molecule miRNA modulators to overcome anti-sense agents' limitations; general enhancers of miRNA maturation; and Argonaute 2 protein and its pharmacokinetic parameters. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and thorough, Drug Target miRNA: Methods and Protocols is a valuable resource for anyone interested in the ever-evolving field of miRNA drug discovery.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

MiRNA Targeting Drugs: The Next Blockbusters?.- Functional Analysis of miRNA using DIANA Tools Online Suite.- Non-Nucleotide Modification of Anti-miRNA Oligonucleotides.- Quantification of Oligonucleotide Association with miRNA-Argonaute Complexes In Vitro.- Determination of Anti-miR Association with miRNA/Argonaute Complexes In Vivo.- Competitive Argonaute-Based RNA Immunoprecipitation for Investigation of Transcriptomic Response to Anti-miR.- Assessing Anti-miR Pharmacology with miRNA Polysome Shift Assay.- Evaluating Synergistic Effects of miR-34a Mimics in Combination with Other Therapeutic Agents in Cultured Non-Small Cell Lung Cancer Cells.- Assessing the Off-Target Effects of miRNA Inhibitors on Innate Immune Toll-Like Receptors.- Design of Multimodal Small Molecules Targeting miRNA Biogenesis: Synthesis and In Vitro Evaluation.- Machine Learning Approaches towards Building Predictive Models for Small MoleculeModulators of miRNA and its Utility in Virtual Screening of Molecular Databases.- Identification of Small Molecule Modulators of MicroRNA by Library Screening.- Rapid Generation of miRNA Inhibitor Leads by Bio-Informatics and Efficient High-Throughput Screening Methods.- Peptide-Based Inhibition of miRNA-Guided Gene Silencing.- Small Molecules Targeting the miRNA Binding Domain of Argonaute 2: From Computer-Aided Molecular Design to RNA Immunoprecipitation.- Surface Plasmon Resonance: A Useful Strategy for the Identification of Small Molecule Argonaute 2 Protein Binders.- Antagonists of the miRNA-Argonaute 2 Protein Complex: Anti-miR-AGOs.- Elucidating Mechanisms of Molecular Recognition between Human Argonaute and miRNA using Computational Approaches.- Kinetic Analysis of Target RNA Binding and Slicing by Argonaute 2 Protein.- Site-Specific Fluorescent Labeling of Argonaute for FRET-Based Bio-Assays.- Single-Molecule Fluorescence Energy Transfer Assays for the Characterization of Reaction Pathways of miRNA-Argonaute Complex.