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This book provides an overview of RNA-based technologies, covering their mechanisms, therapeutic applications, and challenges. Chapters explore therapeutic RNAs, RNA stability, mRNA vaccine formulation, strategies to overcome immunogenicity, delivery hurdles, mRNA-based cancer immunotherapy, CAR and TCR T cell therapies, and RNA interference. Additional chapters examine RNA delivery systems such as lipid nanoparticles, nanotubular structures and extracellular vesicles. Written in the highly successful Methods in Molecular Biology series format, the chapters include brief introductions to the material, lists of necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and a Notes section which highlights tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, RNA Therapeutics: From siRNA to mRNA Innovations aims to be comprehensive guide for researchers in the field.
List of contents
RNA therapeutics: Bridging discovery and clinical implementation.- Molecular mechanisms of innate immune sensing of exogenous RNAs.- Chemical modifications in nucleic acid therapeutic.- Effects of nucleoside modifications on mRNA Translation: Choosing the right modifications.- Generation of lipid nanoparticle mRNA vaccines and evaluation of antigen specific CD8+ T cell responses.- Assessing the immunogenicity of synthetic RNA using blood cells.- Lateral flow immunoassay for rapid and sensitive detection of dsRNA contaminants in in vitro-transcribed mRNA products.- Hydrophobic chromatography purification of linearized Plasmid DNA reduces dsRNA formation during in vitro transcription.- Optimization of In vitro transcription reaction for mRNA production using chromatographic at-line monitoring.- Large scale production of unmodified mRNA for reprogramming human dendritic cells and T cells.- Production of mRNA-loaded dendritic cell cancer vaccines.- Enhancing cancer vaccine efficacy: siRNA-based modulation of immune suppressive factors in dendritic cells.- mRNA-based engineering of viral antigen-specific TCR-T cells.- Engineering mRNA CAR-T cells for cancer immunotherapy.- Generating mRNA encoding anti-HBV designer epigenome modifiers.- Lipid nanoparticles for the delivery of mRN.- Optimization of DOTAP/cholesterol cationic lipid nanoparticles for nucleic acid delivery.- Formulation and characterization of cationic bicelles for siRNA delivery.- Bioinspired extracellular vesicles for enhanced delivery of siRNA to tumours.- Construction of nanotube-shaped mRNA vehicles using self-assembling peptides.- Nanoparticle-mediated siRNA delivery in human epithelial lung cells cultured at the air-liquid interface.- Developing aptamer-targeted mRNA for immunotherapy.- A facile method for assessing cellular stability and co-localization of Cas9 mRNA and sgRNA using confocal microscopy.- siRMSD: A structural parameter to reduce sequence-dependent off-target effects for siRNA design with chemical modifications.- Chemical strategies to enhance antisense strand selection and minimize off-target effect-mediated by siRNAs.- In vivo fermentation production of RNA interference agents.
Summary
Chapters explore therapeutic RNAs, RNA stability, mRNA vaccine formulation, strategies to overcome immunogenicity, delivery hurdles, mRNA-based cancer immunotherapy, CAR and TCR T cell therapies, and RNA interference.
