Full Description
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.
Contents
Tour de Herpes: Cycling through the Life and Biology of HSV-1.- Vaccines for Herpes Simplex: Recent Progress Driven by Viral and Adjuvant Immunology.- Herpes Simplex Virus Growth, Preparation, and Assay.- Engineering HSV-1 Vectors for Gene Therapy.- Preparation of Herpes Simplex Virus Type 1 (HSV-1)-Based Amplicon Vectors.- HSV-1 Amplicon Vectors as Genetic Vaccines.- oHSV Genome Editing by Means of galK Recombineering.- Rescue, Purification, and Characterization of a Recombinant HSV Expressing a Transgenic Protein.- CRISPR/Cas9-Based Genome Editing of HSV.- Latent/Quiescent Herpes Simplex Virus 1 Genome Detection by Fluorescent In Situ Hybridization (FISH).- Oligonucleotide Enrichment of HSV-1 Genomic DNA from Clinical Specimens for use in High-Throughput Sequencing.- HSV Mutant Generation and Dual-Detection Methods for Gaining Insight into Latent/Lytic Cycles In Vivo.- Phenotypic and Genotypic Testing of HSV-1 and HSV-2Resistance to Antivirals.- Using Primary SCG Neuron Cultures to Study Molecular Determinants of HSV-1 Latency and Reactivation.- Characterization of Extracellular HSV-1 Virions by Proteomics.- Analysis and Sorting of Individual HSV-1 Particles by Flow Virometry.- Isolation/Analysis of Extracellular Microvesicles from HSV-1-Infected Cells.- Conformational Change in Herpes Simplex Virus Entry Glycoproteins Detected by Dot Blot.- BioID Combined with Mass Spectrometry to Study Herpesvirus Protein-Protein Interaction Networks.- Preparation of Herpes Simplex Virus Infected Primary Neurons for Transmission Electron Microscopy.- Transmission Immuno-Electron Microscopy of Herpes Simplex Virus-1 Infected Dorsal Root Ganglia Neurons Sectioned in Growth Plane.- Multi-Fluorescence Live Analysis of Herpes Simplex Virus Type-1 Replication.- Expression, Purification, and Crystallization of HSV-1 Glycoproteins for Structure Determination.- Expression, Purification, and Crystallization of Full-Length HSV-1 gB for Structure Determination.- The Use of Microfluidic Neuronal Devices to Study the Anterograde Axonal Transport of Herpes Simplex Virus-1.- A Model of In Vivo HSV-1 DNA Transport using Murine Retinal Ganglion Cells.- The Murine Intravaginal HSV-2 Challenge Model for Investigation of DNA Vaccines.
