The process proceeds in the opposite direction to forward genetic screens of classical genetics… The authors would like to thank Dr. Gerd Hobom (Institut fur Mikro- und Molekularbiologie, Giessen, Germany) for kindly providing pHH21 vector and Dr. Yoshihiro Kawaoka (University of Wisconsin) for providing plasmids for A/WSN/33 (H1N1) reverse genetics. PLoS One 7:e46378, Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Older browsers that do not support HTML5 and the H.264 video codec will still use a Flash-based video player. Virol J 4:102, Song MS, Baek YH, Pascua PN, Kwon HI, Park SJ, Kim EH, Lim GJ, Choi YK (2013) Establishment of Vero cell RNA polymerase I-driven reverse genetics for Influenza A virus and its application for pandemic (H1N1) 2009 influenza virus vaccine production. We use cookies to enhance your experience on our website. Editors: By continuing to use our website or clicking “Continue”, you are agreeing to accept our cookies. Arch Virol 158:2049–2058, Mostafa A, Kanrai P, Ziebuhr J, Pleschka S (2013) Improved dual promotor-driven reverse genetics system for influenza viruses. Over 10 million scientific documents at your fingertips. Please check your Internet connection and reload this page. Reverse genetics is the creation of a virus from a full-length cDNA copy of the viral genome, referred to as an “infectious clone,” and is one of the most powerful genetic tools in modern virology. Springer Nature is developing a new tool to find and evaluate Protocols. Virology 267:310–317, Neumann G, Fujii K, Kino Y, Kawaoka Y (2005) An improved reverse genetics system for influenza A virus generation and its implications for vaccine production. Cite as. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Perez, Daniel R. © 2020 Springer Nature Switzerland AG. Learn more. This volume is a compilation of sixteen chapters that detail reverse genetics protocols. Not logged in All rights reserved, Degranulation: Effects of Triclosan without Use of an Organic Solvent, A Microplate Assay to Assess Chemical Effects on RBL-2H3 Mast Cell, Using Light Sheet Fluorescence Microscopy to Image Zebrafish Eye Development. The JoVE video player is compatible with HTML5 and Adobe Flash. This is a preview of subscription content, Engelhardt OG (2013) Many ways to make an influenza virus: review of influenza virus reverse genetics methods. enable JavaScript in your browser. We may use this info to send you notifications about your account, your institutional access, and/or other related products. Proc Natl Acad Sci U S A 102:16825–16829, Chen H, Ye J, Xu K, Angel M, Shao H, Ferrero A, Sutton T, Perez DR (2012) Partial and full PCR-based reverse genetics strategy for influenza viruses. Although continual improvement in reverse genetics system is being made in different laboratories for the efficient rescue of the influenza virus, the basic concept of synthesizing viral RNA using RNA polymerase I remains the same. CYBER DEAL: 50% off all Springer eBooks | Get this offer! Your access has now expired. Reverse genetics systems have truly transformed our ability to manipulate and study negative-strand RNA … A subscription to JoVE is required to view this content.You will only be able to see the first 20 seconds. Clin Vaccine Immunol 13:395–402, Czudai-Matwich V, Schnare M, Pinkenburg O (2013) A simple and fast system for cloning influenza A virus gene segments into pHW2000- and pCAGGS-based vectors. 2 Reverse genetics systems facilitate investigation of many aspects of the life cycle and pathogenesis of viruses. Authoritative and practical, Reverse Genetics of RNA Viruses: Methods and Protocols aims to ensure successful results in the further study of this vital field. J Virol Methods 193:603–610, Massin P, Rodrigues P, Marasescu M, van der Werf S, Naffakh N (2005) Cloning of the chicken RNA polymerase I promoter and use for reverse genetics of influenza A viruses in avian cells. Proc Natl Acad Sci U S A 93:11354–11358, Lamb RA, Krug RM (2001) Orthomyxoviridae: the viruses and their replication. Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins Article doi: 10.3791/61039 April 17th, 2020 • Asha A. Philip 1, Jin Dai 1, Sarah P. Katen 1, John T. … JavaScript is currently disabled, this site works much better if you Copyright © 2020 MyJoVE Corporation. Reverse genetics is the process of generating an RNA virus from a cDNA copy. Vaccine 22:3175–3181, Lee CW, Lee YJ, Senne DA, Suarez DL (2006) Pathogenic potential of North American H7N2 avian influenza virus: a mutagenesis study using reverse genetics. Vaccine 20:3165–3170, Wang L, Lee CW (2009) Sequencing and mutational analysis of the non-coding regions of influenza A virus. If you want more info regarding data storage, please contact gdpr@jove.com. 50.62.23.88. We use/store this info to ensure you have proper access and that your account is secure. Reverse Genetics for Mammalian Orthoreovirus, Development and Characterization of an Infectious cDNA Clone of Equine Arteritis Virus, Reverse Genetics for Porcine Reproductive and Respiratory Syndrome Virus, Efficient Reverse Genetic Systems for Rapid Genetic Manipulation of Emergent and Preemergent Infectious Coronaviruses, Reverse Genetics System for the Avian Coronavirus Infectious Bronchitis Virus, BAC-Based Recovery of Recombinant Respiratory Syncytial Virus (RSV), Recovery of a Paramyxovirus, the Human Metapneumovirus, from Cloned cDNA, Reverse Genetics of Newcastle Disease Virus, Rapid Reverse Genetics Systems for Rhabdoviruses: From Forward to Reverse and Back Again, Rescue of Infectious Salmon Anemia Virus (ISAV) from Cloned cDNA, Plasmid-Based Reverse Genetics of Influenza A Virus.