Hepatitis B Virus Methods and Protocols

This detailed new edition collects state-of-the-art methods in the fields of hepatitis B virus (HBV) molecular biology and immunology. The book features techniques for cell culture, epigenetic regulation of cccDNA transcription, epitranscriptomic modification of HBV RNA, HBV pregenomic RNA and subviral particle morphogenesis, and much more. 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, Hepatitis B Virus: Methods and Protocols, Second Edition serves as an ideal guide for researchers pursuing a cure for this chronic disease.

Stem Cell-Derived Hepatocyte-Like Cells for Hepatitis B Virus Infection.- Primary Duck Hepatocyte Culture and Duck Hepatitis B Virus Infection Model.- Hepatitis B Virus Covalently Closed Circular DNA Chromatin Immunoprecipitation Assay.- Analysis of HBV cccDNA Minichromosome Accessibility by MNase-qPCR and High Throughput Sequencing.- Circular Chromosome Conformation Capture (4C-Seq) Analysis of HBV-Host Chromosome DNA Interactome.- Analysis of N6-Methyladenosine Modification of HBV RNA by Methylated RNA Immunoprecipitation.- Structural Analysis of the Hepatitis B Virus RNA Encapsidation Signal ? by Selective 2'-Hydroxyl Acylation Analyzed by Primer Extension (SHAPE).- A Particle Gel Assay for Detection of Intracellular Hepatitis B Virus Subviral Particles in Cultured Cells.- Detection of Hepatitis B Virus Covalently Closed Circular DNA and Intermediates in Its Formation.- Rapid and Reliable Protein-Free HBV DNA Extraction and Sensitive Branched DNA Southern Blot Assay.- A Fluorescent In Situ Hybridization (FISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Cell Culture.- A Chromogenic In Situ Hybridization (CISH) Assay for Detection of HBV RNA, DNA, and cccDNA in Liver Tissue.- Detection and Quantification of HBV Transcripts by Full-Length 5'RACE-PCR.- One-Step Reverse Transcriptase qPCR Method for Serum Hepatitis B Virus RNA Quantification.- Method for Quantitative HDV RNA Detection: I, Manual Workflow (Serum and Liver Tissue) and II, Fully Automated High Throughput Workflow for Diagnostic Use.- The Cre/loxP-Based Recombinant HBV cccDNA System In Vitro and In Vivo.- Modeling HBV Infection and Therapy in Immunodeficient NOD-Rag1-/-IL2RgammaC-null (NRG) Fumarylacetoacetate Hydrolase (FAH) Knockout Mice with Human Chimeric Liver.- Production and Purification of Adeno-Associated Viral Vectors for the Development of Immune-Competent Mouse Models of Persistent Hepatitis B Virus Replication.- Peptide-Loaded HLA Class I Tetramer-Associated Magnetic Bead-Based Enrichment of HBV-Specific CD8+ T Cells.- Optimized Ex Vivo Fluorospot Assay to Measure Multi-Functional HBV-Specific T Cell Frequencies in Chronic Hepatitis B Patients.- Ex Vivo Fluorescent Labeling of HBV-Specific B Cells in Chronic Hepatitis B Patients.- In Vitro Enzymatic and Cell Culture-Based Assays for Measuring Activity of HBV Ribonuclease H Inhibitors.