Antiviral Methods And Protocols

This latest addition to the Methods in Molecular Medicine series, Anti- ral Methods and Protocols, is opportune because there is an increasing int- est in discovering compounds that are effective against both chronic and acute viral infections. A number of the methods described in the volume are unp- lished and their inclusion indicates the speed at which this field is moving. This volume is not a review but each chapter contains methods validated by the experts who have spent time in developing the protocols. The hallmark of this series is the comprehensive way in which the me- ods are described, which includes a list of all the reagents needed for each protocol. Of importance is the section on tips and pitfalls that the authors have discovered while developing their protocols. The manual itself is designed to be used by researchers in universities and industry who are familiar with a range of biological techniques but who want to set up quickly a novel assay system. We encourage a dialog between readers and authors, which may also result in useful collaborations.

Although antiviral drugs have been successfully developed for some viral diseases, there remains a clear, unmet medical need to develop novel antiviral agents for the control and management of many viruses that currently have no or limited treatment options as well as a need to overcome the limitations associated with the existing antiviral drugs, such as adverse effects and emergence of drug-resistant mutations. The second edition of Antiviral Methods and Protocols features: All chapters are new and written by experts in the field, reflecting the major recent technical advances in antiviral research and discovery. This edition focuses on many important human viruses, such as human immunodeficiency virus type 1 (HIV-1), hepatitis viruses (hepatitis B and C viruses), herpes viruses, human respiratory syncytial virus (RSV), and influenza virus, while also featuring some important emerging viruses, such as dengue virus, West Nile virus, and chikungunya virus. As a volume in the highly successful Methods in Molecular Biology series, 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. Comprehensive and cutting-edge, Antiviral Methods and Protocols, Second Edition will serve as an excellent laboratory reference for pharmaceutical and academic biologists, medicinal chemists, and pharmacologists as well as for virologists in the field of antiviral research and drug discovery.

Viruses and RNAi share an intricate relationship at many levels. RNAi is an important antiviral defense mechanism in plants and invertebrates, microRNAs – of viral or cellular origin – affect many aspects of virus biology, and replication of many, if not all, mammalian viruses can be suppressed by RNAi. Antiviral RNAi: Concepts, Methods, and Applications provides a collection of protocols for the analysis of viral small RNAs and natural antiviral RNAi responses as well as for the development and optimization of RNAi-based antiviral drugs. As RNAi is a central regulatory mechanism in the cell, the methods in this volume can also be applied out of the context of a virus infection. Divided into five convenient parts, this detailed volume reviews important basic concepts in the field of antiviral RNAi, provides experimental and bio-informatic tools for the analysis of small silencing RNAs, covers methods to biochemically dissect RNAi-based antiviral defense and viral counter-defense mechanisms, describes methods for the design, expression, and delivery of therapeutic antiviral siRNAs, and finally presents genome-wide RNAi approaches for the identification of factors involved in virus replication. Written in the highly successful Methods in Molecular BiologyTM series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Antiviral RNAi: Concepts, Methods, and Applications serves as an ideal guide for both novice and experienced researchers alike striving to dissect the role of RNAi in the viral life cycle or to further boost the development of novel therapeutics and experimental tools based on RNAi technology.

This book explores methods to study the complex and evolving interplay between a virus and its host that range from model systems to the detection of chemical molecules. The collection starts with the application of humanized mice and zebrafish as model organisms to study virus-host interactions and induction of innate immune responses. Subsequent chapters outline diverse methods to detect small interfering RNAs, microRNAs, and virus-derived dsRNA from a variety of cells, tissues, and organisms, as well as to interrogating the cytosolic RNA and DNA sensing pathways, including using RNA PAMPs as molecular tools, purification of cGAMP from virus particles and infected cells, and mechanisms to visualize the subcellular localization and activation of the adaptor proteins MAVS and STING. Cutting-edge methods, including high-throughput and genome-wide CRISPR/Cas9 screens, chromosome conformation capture, and whole-exome sequencing, are described to identify novel mediators, pathways, and variants underlying host susceptibility. Given the importance of studying these pathways and players under physiologic conditions, methods describing the isolation of primary mouse sensory neurons and group 2 innate lymphoid cells are also provided. Finally, this collection comes full circle back to the whole organism level and concludes with epidemiological methods to investigate virus-host interactions and the induction of innate immunity. Written for the Methods in Molecular Biology series, 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. Authoritative and practical, Innate Antiviral Immunity: Methods and Protocols spans a diverse array of approaches to study and elucidate the intricacies of this vital area of study. The chapter 'Morphological Separation of Clustered Nuclei in Histological Images' is published open access under a CC BY 4.0 license at link.springer.com.

This second edition volume expands on the previous edition with a discussion of new and updated methods used to study the Herpes Simplex Virus (HSV), along with a look at the latest developing technologies such as next generation sequencing, CRISPR/Cas9 engineering, and the use of BioID to identify protein-protein interactions. Chapters cover topics such as the biology, life cycle, and current state of antiviral and vaccine development for HSV-1; protocols on growing viruses in cell culture and manipulating viral DNA; design and application of HSV-1 vectors for cancer- and gene-therapy; and structural analyses, microscopy, proteomics, and testing of antivirals. 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. Cutting-edge and comprehensive, Herpes Simplex Virus: Methods and Protocols, Second Edition is a valuable resource for immunologists, and molecular and cell biologists. This book will also be useful for researchers who wish to initiate molecular and/or cellular-based approaches to study HSV. .

This book provides researchers with widely used techniques for the study of virology, focusing on molecular biology and imaging to encourage mechanistic investigation of virus-host interactions. Chapters detail a broad range of methods from diagnosis, virus propagation, proteomics, haploid screening, lentiviral screening, virus entry, single molecule RNA imaging, correlative light and electron microscopy (CLEM), EM, light-sheet microscopy, biochemistry, viral transcription, physiological infection models, animal models, in vivo imaging, antigenic evolution, immunology to mathematical modelling. Reviews cover general influenza, clinical trials, both sides of the gain-of-function debate, and computational modelling. 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. Cutting-edge and thorough, Influenza Virus: Methods and Protocols aims to motivate experienced researchers and newcomers in the field and improve our overall understanding of influenza.

Reports of influenza-like illnesses date back to the Middle Ages, and outbreaks of influenza likely afflicted humans long before that. Over the last half century, influenza virus research has led to the development of two classes of antivirals – ion channel and neuraminidase inhibitors. Recently, a method of the artificial generation of an influenza virus was established. This system has been instrumental in the development of novel influenza vaccines and in the understanding of viral pathogenicity and the functions of viral proteins. Influenza Virus: Methods and Protocols summarizes the current techniques that have made this progress possible, ranging from protocols for virus isolation, growth, and subtyping to procedures for the efficient generation of any influenza virus. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Influenza Virus: Methods and Protocols seeks to serve both professionals and novices with the techniques used in numerous laboratories around the world that are, thus, the building blocks that underpin almost all influenza virus research.