This detailed volume explores a variety of techniques for studying antiviral innate immunity and the type I interferon (IFN-I) signaling pathway. Beginning with guidelines to identify novel proteins from the host or viruses that regulate the antiviral IFN-I signaling pathway, the book continues with common and cutting-edge approaches for identifying novel regulatory factors, posttranslational modifications, as well as click chemistry to detect protein modifications. Other chapters delve into topics such as cytosolic RNA and DNA sensors, magnetic beads, cryo-electron microscopy, zebrafish, humanized mice, and organoids as model organisms to study host-virus interactions, and epidemiological approaches. Written for 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Antiviral Innate Immunity serves as an ideal guide for researchers working in this vital area of the virology and innate immunity fields.

Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts

Inhalt

A Guideline Strategy for Identifying Genes/Proteins Regulating Antiviral Innate Immunity.- A Guideline Strategy for Identifying a Viral Gene/Protein Evading Antiviral Innate Immunity.- Luciferase Reporter Assay for Determining the Signaling Activity of Interferons.- Identifying an Abnormal Phosphorylated Adaptor by Viral Kinase Using Mass Spectrometry.- Identifying a Ubiquitinated Adaptor Protein by a Viral E3 Ligase Through Co-Immunoprecipitation.- CRISPR-Mediated Viral Gene Knockout to Investigate Viral Evasion of Antiviral Innate Immunity.- CRISPR-Mediated Library Screening of Gene-Knockout Cell Lines for Investigating Antiviral Innate Immunity.- CRISPR-Mediated Construction of Gene-Knockout Mice for Investigating Antiviral Innate Immunity.- Click Chemistry in Detecting Protein Modification.- Transcriptomics in the Study of Antiviral Innate Immunity.- Application of Proteomics Technology Based on LC-MS Combined with Western Blotting and Co-IP in Antiviral Innate Immunity.- Identifying RNA Sensors in Antiviral Innate Immunity.- DNA PAMPs as Molecular Tools for the cGAS-STING Signaling Pathways.- Luciferase Reporter Systems in Investigating Interferon Antiviral Innate Immunity.- Investigation of Protein Lysine Acetylation in Antiviral Innate Immunity.- In Vitro Cleavage Assay to Characterize DENV NS2B3 Antagonism of cGAS.- Phagosome Isolation with Magnetic Beads and Purification of Toll-Like Receptor (TLR) Complexes in Phagosomes.- Cryo-Electron Microscopy in the Study of Antiviral Innate Immunity.- Epidemiological Study in Antiviral Innate Immunity.- The Application of Organoids in the Study of Antiviral Innate Immunity.- Yeast Two-Hybrid Assay for Investigating Antiviral Innate Immunity.- Zebrash as a Model for Investigating Antiviral Innate Immunity.- In Vitro and In Vivo Evaluation of Virus-Induced Innate Immunity in Mice.- Identifying a Gene Deficiency in the Antiviral Innate Immune Signaling Pathway.- Protein Kinase R (PKR) as a Novel dsRNA Sensor in Antiviral Innate Immunity.