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This book provides a collection of comprehensive, up-to-date, and broadly applicable guides to the research and development fields of oligonucleotide (ON) therapeutics. Covering topics from the study of antisense and anti-gene effects to oligonucleotides in the context of drug discovery and development, the volume explores a wide-ranging and useful spectrum of methods and protocols needed to take full advantage of therapeutic applications involving ONs. Written for the highly successful Methods in Molecular Biology series, 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.
Authoritative and practical, Oligonucleotide-Based Therapies: Methods and Protocols aims to be a great aid in the laboratory as well as an ideal reference guide when designing antisense and anti-gene oligonucleotides for therapeutic applications.
List of contents
Chemical Development of Therapeutic Oligonucleotides.- Tissue-Specific Delivery of Oligonucleotides.- Designing siRNA and Evaluating Its Effect on RNA Targets Using qPCR and Western Blot.- Antisense Oligonucleotides for Splice Modulation: Assessing Splice Switching Efficacy.- Assessing Oligonucleotide Binding to Double-Strand DNA.- Modelling and Simulation of Oligonucleotide Hybrids: Outlining a Strategy.- Characterization of Peptide/Oligonucleotide Complexes Using Electron Microscopy, Dynamic Light Scattering, and Protease Resistance Assay.- Click-Shielded and Targeted Lipopolyplexes.- Attachment of Peptides to Oligonucleotides on Solid Support Using Copper(I)-Catalyzed Huisgen 1,3-Dipolar Cycloaddition.- Characterizing Oligonucleotide Uptake in Cultured Cells: A Case Study Using AS1411 Aptamer.- Measuring the Action of Oligonucleotide Therapeutics in the Lung at the Cell Type-Specific Level by Tissue Disruption and Cell Sorting (TDCS).- Delivery of Antisense Oligonucleotides Mediatedby a Hydrogel System: In Vitro and In Vivo Application in the Context of Spinal Cord Injury.- Evaluation of Cell-Penetrating Peptide Delivery of Antisense Oligonucleotides for Therapeutic Efficacy in Spinal Muscular Atrophy.- Injection and Infusion of Compounds to the Central Nervous System.- In Vitro Assessment of the Hepatotoxicity Potential of Therapeutic Oligonucleotides.- Identifying Suitable Target Regions and Analyzing Off-Target Effects.- RACE-SEQ and Population-Wide Polymorphism Susceptibility Testing for Endonucleolytically Active, RNA-Targeting Therapeutics.- Studying the Biotransformation of Phosphorothioate-Containing Oligonucleotide Drugs by LC-MS.- ADME: Assessing Pharmacokinetic-Pharmacodynamic Parameters of Oligonucleotides.
