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 necessarymaterials 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.