Synthetic mRNA
Description
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.
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Content
Introduction into Cells, and Physiological ConsequencesRobert E. Rhoads
Part
II Synthesis of mRNA2.
Synthetic
Capped mRNAs for Cap-Specific Photo-Crosslinking ExperimentsJoanna Kowalska, Franck Martin, and Jacek Jemielity3. Enzymatic Modification of 5'-Capped
RNA and Subsequent Labeling by Click ChemistryJosephin M. Holstein, Daniela Stummer, and
Andrea Rentmeister4. Preparation of Functional,
Fluorescently Labeled mRNA Capped with Anthraniloyl-m7GpppGArtem V. Domashevskiy, David J.
Rodriguez, Dilantha Gunawardana, and Dixie J. Goss5. Intronless b-Globin
Reporter: A Tool for Studying Nuclear
RNA Stability ElementsJessica A. Brown and Joan A. Steitz6.
Synthetic mRNA with Superior Properties that Mimics
the Intracellular Fates of Natural Histone mRNAWei Su, Michael K. Slevin, William F. Marzluff, and Robert E.
Rhoads7. Engineering WT1-Encoding mRNA to
Increase Translational Efficiency in Dendritic CellsDaphné Benteyn, Carlo Heirman, Kris
Thielemans, and Aude BonehillPart
III Introduction of Synthetic mRNA into Cells8. Electroporation of Alphavirus RNA
Translational Reporters into Fibroblastic and Myeloid Cells as a Tool to Study
the Innate Immune SystemChristina L. Gardner, Derek W. Trobaugh,
Kate D. Ryman, and William B. Klimstra9. GMP-Grade mRNA Electroporation of
Dendritic Cells for Clinical UseJudith Derdelinckx, Zwi N. Berneman, and Nathalie
Cools10. Large Scale mRNA Transfection of
Dendritic Cells by Electroporation in Continuous Flow SystemsDavid Selmeczi, Thomas Steen Hansen,
Özcan Met, Inge Marie Svane, and Niels B. Larsen11. FLT3 Ligand as a Molecular Adjuvant
for Naked RNA VaccinesSebastian Kreiter, Mustafa Diken,
Abderraouf Selmi, Jutta Petschenka, Özlem Türeci, and Ugur Sahin12. Transfecting Human Monocytes with
RNAJens Dannull and Smita K. Nair13.< In Vitro Synthesis, Delivery and
Bioavailability of Exogenous mRNA in Gene Transfer Mediated by PiggyBac TranspositionSolenne Bire, Nicole Ishac, and Florence
Rouleux-Bonnin14. Transfection of Human Keratinocytes
with Nucleoside-Modified mRNA Encoding CPD-Photolyase to Repair DNA DamageGábor Boros, Katalin Karikó, Hiromi
Muramatsu, Edit Miko, Eszter Emri, Csaba Hegedus, Gabriella Emri, and Éva
RemenyikPart
IV Alteration of Cellular Function with Synthetic mRNA15. Delivery of Synthetic mRNA Encoding
FOXP3 Antigen into Dendritic Cells for Inflammatory Breast Cancer ImmunotherapyGayathri R. Devi and Sritama Nath16. Immune Monitoring using mRNA-Transfected
Dendritic CellsTroels Holz Borch, Inge Marie Svane, and Özcan
Met17. Transfection of Tumor-Infiltrating
T Cells with mRNA Encoding CXCR2Manja Idorn, Per thor Straten, Inge Marie
Svane, and Özcan Met18. mRNA Electroporation of Dendritic
Cells with WT1, Survivin and TriMix (A Mixture of caTLR4, CD40L, and CD70)An Coosemans, Sandra Tuyaerts, Kim
Morias, Jurgen Corthals, Carlo Heirman, Kris Thielemans, Stefaan W. Van Gool,
Ignace Vergote, and Frédéric Amant19. Redirecting T Cell Specificity
Using T Cell Receptor Messenger RNA ElectroporationSarene Koh, Noriko Shimasaki, and Antonio
Bertoletti20. Measuring Hematocrit in Mice Injected
with In Vitro-Transcribed Erythropoietin mRNAAzita Josefine Mahiny and Katalin Karikó21. Genetic Modification of Human
Pancreatic Progenitor Cells through Modified mRNASong Lu, Christie C. Chow, Junwei Zhou,
Po Sing Leung, Stephen K. Tsui, and Kathy O. Lui