Full Description
This volume serves as a reference for the dissemination of advances made in the study of Hepatitis B Virus (HBV). Hepatitis B Virus: Methods and Protocols details protocols and techniques ranging from cell culture studies to in vivo and clinical immunology. The chapters in this book discuss treatments of in vitro infection systems, analysis and quantification of cccDNA and its mutations; in vitro polymerase activity assays; cellular trafficking of core proteins; intracellular calcium metabolism; detection, cloning, and sequencing of HBV markers; and new strategies aimed at exploiting new mechanisms for drug discovery. The book also covers classical methods for resolution of extracellular viral particles by native gel electrophoresis, and methods for detecting HBV antigens in drug discovery. 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, Hepatitis B Virus: Methods and Protocols is a valuable tool for researchers to use toward their advanced studies in HBV.
Contents
NTCP-Reconstituted In Vitro HBV Infection System.- Hepatitis B Virus Infection of HepaRG Cells, HepaRG-hNTCP Cells and Primary Human Hepatocytes.- Live Cell Imaging Confocal Microscopy Analysis of HBV Myr-PreS1 Peptide Binding and Uptake in NTCP-GFP Expressing HepG2 Cells.- Intracytoplasmic Transport of Hepatitis B Virus Capsids.- A Homokaryon Assay for Nucleocytoplasmic Shuttling Activity of HBV Core Protein.- Analyses of HBV cccDNA Quantification and Modification.- Detection of HBV cccDNA Methylation from Clinical Samples by Bisulfite Sequencing and Methylation Specific PCR.- A T7 Endonuclease I Assay to Detect TALEN-Mediated Mutation of HBV cccDNA.- Detection of Hepatocyte Clones Containing Integrated Hepatitis B Virus DNA using Inverse Nested PCR.- Highly Sensitive Detection of HBV RNA in Liver Tissue by In Situ Hybridization.- Immunofluorescent Staining for the Detection of the Hepatitis B Core Antigen in Frozen Liver Sections of Human Liver Chimeric Mice.- Measuring Changes in Cytosolic Calcium Levels in HBV- and HBx-Expressing Cultured Primary Hepatocytes.- In Vitro Assays for RNA Binding and Protein Priming of Hepatitis B Virus Polymerase.- In Vitro Enzymatic and Cell Culture-Based Assays for Measuring Activity of HBV RNaseH Inhibitors.- Detection of Hepatitis B Virus Particles Released from Cultured Cells by Particle Gel Assay.- Microtiter-Format Assays for HBV Antigen Quantitation in Non-Clinical Applications.- Deep Sequencing of the Hepatitis B Virus Genome: Analysis of Multiple Samples by Implementation of the Illumina Platform.- Generation of Replication Competent Hepatitis B Virus Genome from Blood Samples for Functional Characterization.- Hydrodynamic HBV Transfection Mouse Model.- An ELISPOT-Based Assay to Measure HBV-Specific CD8+ T Cell Responses in Immunocompetent Mice.- Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells andKupffer Cells.- Partial Hepatectomy and Castration of HBV Transgenic Mice.- Studying HBV Infection and Therapy in Immune Deficient NOD-Rag1-/-IL2RgammaC-null (NRG) Fumarylacetoacetate Hydrolase (Fah) Knockout Mice Transplanted with Human Hepatocytes.- Measurement of Antiviral Effect and Innate Immune Response during Treatment of Primary Woodchuck Hepatocytes.
