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by Robert E. Rhoads (Editor)
Part I Introduction
1. Synthetic mRNA: Production, Introduction into Cells, and Physiological ConsequencesRobert E. Rhoads Part II Synthesis of mRNA 2. Synthetic Capped mRNAs for Cap-Specific Photo-Crosslinking Experiments
Joanna Kowalska, Franck Martin, and Jacek Jemielity
3. Enzymatic Modification of 5'-Capped RNA and Subsequent Labeling by Click Chemistry
Josephin M. Holstein, Daniela Stummer, and Andrea Rentmeister
4. Preparation of Functional, Fluorescently Labeled mRNA Capped with Anthraniloyl-m7GpppG
Artem V. Domashevskiy, David J. Rodriguez, Dilantha Gunawardana, and Dixie J. Goss
5. Intronless b-Globin Reporter: A Tool for Studying Nuclear RNA Stability Elements
Jessica A. Brown and Joan A. Steitz
6. Synthetic mRNA with Superior Properties that Mimics the Intracellular Fates of Natural Histone mRNA
Wei Su, Michael K. Slevin, William F. Marzluff, and Robert E. Rhoads
7. Engineering WT1-Encoding mRNA to Increase Translational Efficiency in Dendritic Cells
Daphné Benteyn, Carlo Heirman, Kris Thielemans, and Aude Bonehill Part III Introduction of Synthetic mRNA into Cells 8. Electroporation of Alphavirus RNA Translational Reporters into Fibroblastic and Myeloid Cells as a Tool to Study the Innate Immune System
Christina L. Gardner, Derek W. Trobaugh, Kate D. Ryman, and William B. Klimstra
9. GMP-Grade mRNA Electroporation of Dendritic Cells for Clinical Use
Judith Derdelinckx, Zwi N. Berneman, and Nathalie Cools
10. Large Scale mRNA Transfection of Dendritic Cells by Electroporation in Continuous Flow Systems
David Selmeczi, Thomas Steen Hansen, Özcan Met, Inge Marie Svane, and Niels B. Larsen
11. FLT3 Ligand as a Molecular Adjuvant for Naked RNA Vaccines
Sebastian Kreiter, Mustafa Diken, Abderraouf Selmi, Jutta Petschenka, Özlem Türeci, and Ugur Sahin
12. Transfecting Human Monocytes with RNA
Jens Dannull and Smita K. Nair
13.In Vitro Synthesis, Delivery and Bioavailability of Exogenous mRNA in Gene Transfer Mediated by PiggyBac Transposition
Solenne Bire, Nicole Ishac, and Florence Rouleux-Bonnin
14. Transfection of Human Keratinocytes with Nucleoside-Modified mRNA Encoding CPD-Photolyase to Repair DNA Damage
Gábor Boros, Katalin Karikó, Hiromi Muramatsu, Edit Miko, Eszter Emri, Csaba Hegedűs, Gabriella Emri, and Éva Remenyik Part IV Alteration of Cellular Function with Synthetic mRNA
15. Delivery of Synthetic mRNA Encoding FOXP3 Antigen into Dendritic Cells for Inflammatory Breast Cancer Immunotherapy
Gayathri R. Devi and Sritama Nath
16. Immune Monitoring using mRNA-Transfected Dendritic Cells
Troels Holz Borch, Inge Marie Svane, and Özcan Met
17. Transfection of Tumor-Infiltrating T Cells with mRNA Encoding CXCR2
Manja Idorn, Per thor Straten, Inge Marie Svane, and Özcan Met
18. 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 Aman
Back Jacket
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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