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.