Synthetic mRNA

This volume§presents detailed laboratory protocols for in§vitro synthesis of mRNA with favorable properties, its introduction into§cells by a variety of techniques, and the measurement of physiological and§clinical consequences such as protein replacement and cancer immunotherapy. Synthetic§techniques are described for structural features in mRNA that provide investigational§tools such as fluorescence emission, click chemistry, photo-chemical§crosslinking, and that produce mRNA with increased stability in the cell, increased translational efficiency, and reduced§activation of the innate immune response. Protocols are described for clinical§applications such as large-scale transfection of dendritic cells, production of§GMP-grade mRNA, redirecting T cell specificity, and use of molecular§adjuvants for RNA vaccines. 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§laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.§§Synthetic§mRNA: Production, Introduction into Cells, and Physiological Consequences is a valuable and cutting-edge resource§for both laboratory investigators and clinicians interested in this powerful§and rapidly evolving technology.§§