Full Description
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.
Contents
Understanding Influenza.- Clinical Diagnosis of Influenza.- Influenza A Virus Genetic Tools - From Clinical Sample to Molecular Clone.- Propagation and Titration of Influenza Viruses.- Purification and Proteomics of Influenza Virions.- Haploid Screening for the Identification of Host Factors in Virus Infection.- Phenotypic Lentivirus Screens to Identify Antiviral Single Domain Antibodies.- Deciphering Virus Entry with Fluorescently-Labeled Viral Particles.- Quantitative RT-PCR Analysis of Influenza Virus Endocytic Escape.- Single Molecule Sensitivity RNA FISH Analysis of Influenza Virus Genome Trafficking.- Correlative Light and Electron Microscopy (CLEM) and 3D Methods to Study Architecture of Virus-Infected Cells.- Correlative Light and Electron Microscopy (CLEM) of Influenza Virus Entry and Budding.- Influenza Virus-Liposome Fusion Studies using Fluorescence Dequenching and Cryo-Electron Tomography.- Metal-Tagging Transmission Electron Microscopy and Immunogold Labelling on Tokuyasu Cryosections to Image Influenza A Virus Ribonucleoprotein Transport and Packaging.- Live Imaging of Influenza Viral Ribonucleoproteins using Light-Sheet Microscopy.- Purification of Unanchored Polyubiquitin Chains from Influenza Virions.- Measurement of Influenza Virus Polymerase Activity.- In vitro models to study influenza virus and Staphylococcus aureus super-infection on a molecular level.- Infection of Cultured Mammalian Cells with Aerosolized Influenza Virus.- Animal Models in Influenza Research.- Measuring Influenza Virus Infection using Bioluminescent Reporter Viruses for in vivo Imaging and in vitro Replication Assays.- Selection of Antigenically Advanced Variants of Influenza Viruses.- Assessment of Influenza Virus Hemagglutinin Stalk-specific Antibody Responses.- Analyses of Cellular Immune Responses from Ferrets Following Influenza Virus Infection.- Parameter Estimation in Mathematical Models of Viral Infections using R.- Software for Characterizing the Antigenic and Genetic Evolution of Human Influenza Viruses.- Clinical Trials of Influenza Vaccines - Special Challenges.- The Silver Lining in Gain-of-Function Experiments with Pathogens of Pandemic Potential.- Why Do Exceptionally Dangerous Gain-of-Function Experiments in Influenza?.- How Computational Models Enable Mechanistic Insights into Virus Infection.
