High-Throughput RNAi Screening Methods and Protocols
High-throughput RNAi screening remains one of the most widely used technologies to perform target identification and validation studies in an unbiased manner. These assays are equally important for research and development across academic, biotech, and ph
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David O. Azorsa Shilpi Arora Editors
High-Throughput RNAi Screening Methods and Protocols
METHODS
IN
MOLECULAR BIOLOGY
Series Editor John M. Walker School of Life and Medical Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
For further volumes: http://www.springer.com/series/7651
High-Throughput RNAi Screening Methods and Protocols Editors
David O. Azorsa Institute of Molecular Medicine, Phoenix Children’s Hospital, Phoenix, Arizona, USA
Shilpi Arora Constellation Pharmaceuticals, Cambridge, Massachusetts, USA
Editors David O. Azorsa Institute of Molecular Medicine Phoenix Children’s Hospital Phoenix, Arizona USA
Shilpi Arora Constellation Pharmaceuticals Cambridge, Massachusetts USA
ISSN 1064-3745 ISSN 1940-6029 (electronic) Methods in Molecular Biology ISBN 978-1-4939-6335-5 ISBN 978-1-4939-6337-9 (eBook) DOI 10.1007/978-1-4939-6337-9 Library of Congress Control Number: 2016945160 © 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. 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 Ever since the discovery of small interfering RNA-mediated transcriptional inhibition by Craig Mello and Andrew Fire, there have been great advances in manipulating this cellular mechanism to achieve targeted gene silencing. Short interfering RNA (siRNA) and short hairpin RNA (shRNA) are commonly used for gene silencing and validation of gene function. Both can be implemented using standard techniques of transferring nucleic acids into cells. With the completion of the human genome project and other advances in genomics, nucleic acid sequences that target specific gene transcripts have become available. In addition, both commercial and academic institutions have made advances in chemical synthesis of double-stranded RNA and the development of large shRNA libraries. Through the use of these libraries
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