Synthetic mRNA Production, Introduction Into Cells, and Physiologica
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
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Robert E. Rhoads Editor
Synthetic mRNA Production, Introduction Into Cells, and Physiological Consequences
METHODS
IN
MOLECULAR BIOLOGY
Series Editor John M. Walker School of Life and Medical Sciences University of Hertfordshire Hatfield, Hertfordshire, UK
For further volumes: http://www.springer.com/series/7651
Synthetic mRNA Production, Introduction Into Cells, and Physiological Consequences
Edited by
Robert E. Rhoads Department of Biochemistry and Molecular Biology Louisiana State University Health Sciences Center Shreveport, LA, USA
Editor Robert E. Rhoads Department of Biochemistry and Molecular Biology Louisiana State University Health Sciences Center Shreveport, LA, USA
ISSN 1064-3745 ISSN 1940-6029 (electronic) Methods in Molecular Biology ISBN 978-1-4939-3623-6 ISBN 978-1-4939-3625-0 (eBook) DOI 10.1007/978-1-4939-3625-0 Library of Congress Control Number: 2016936683 © Springer Science+Business Media New York 2016 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. Cover illustration: Bioluminescence image of a living mouse after intranodal injection with synthetic mRNA encoding firefly luciferase (as described by Kreiter et al. in Chapter 11) Printed on acid-free paper This Humana Press imprint is published by Springer Nature The registered company is Springer Science+Business Media LLC New York
Preface The dream of altering genetic expression to alleviate human disease states—gene therapy— has existed from the dawn of the molecular biology era. Early attempts were made with both DNA and RNA as carriers of genetic information, but the inherent instability of RNA compared with DNA attracted by far the larger share of researchers, vectors, protocols, and success stories. Yet DNA, regardless of whether it is introduced by viral or nonviral modalities, carries the potential of integration into the host genome at unintended sites, which can lead to unwelcome and permanent consequences for the patient. Also, there are some outcomes of gene therapy that are best ac
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