This volumepresents detailed laboratory protocols for invitro synthesis of mRNA with favorable properties, its introduction intocells by a variety of techniques, and the measurement of physiological andclinical consequences such as protein replacement and cancer immunotherapy. Synthetictechniques are described for structural features in mRNA that provide investigationaltools such as fluorescence emission, click chemistry, photo-chemicalcrosslinking, and that produce mRNA with increased stability in the cell, increased translational efficiency, and reducedactivation of the innate immune response. Protocols are described for clinicalapplications such as large-scale transfection of dendritic cells, production ofGMP-grade mRNA, redirecting T cell specificity, and use of molecularadjuvants for RNA vaccines. Writtenin the highly successful Methods inMolecular Biology series format, chapters include introductions to theirrespective topics, lists of the necessarymaterials and reagents, step-by-steplaboratory protocols, and tips on troubleshooting and avoiding known pitfalls.SyntheticmRNA: Production, Introduction into Cells, and Physiological Consequences is a valuable and cutting-edge resourcefor both laboratory investigators and clinicians interested in this powerfuland rapidly evolving technology.
