Main 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:

Preface...
Table of Contents...
Contributing Authors...

1. Tour de Herpes: Cycling through the Life and Biology of HSV-1
Christopher E. Denes, Roger D. Everett, and Russell J. Diefenbach

2. Vaccines for Herpes Simplex: Recent Progress Driven by Viral and Adjuvant Immunology
Kerrie J. Sandgren, Naomi R. Truong, Jacinta B. Smith, Kirstie Bertram, and Anthony L. Cunningham

3. Herpes Simplex Virus Growth, Preparation, and Assay
Sereina O. Sutter, Peggy Marconi, and Anita F. Meier

4. Engineering HSV-1 Vectors for Gene Therapy
William F. Goins, Shaohua Huang, Bonnie Hall, Marco Marzulli, Justus B. Cohen, and Joseph C. Glorioso

5. Preparation of Herpes Simplex Virus Type 1 (HSV-1)-Based Amplicon Vectors
Cornel Fraefel and Alberto L. Epstein

6. HSV-1 Amplicon Vectors as Genetic Vaccines
Anita F. Meier and Andrea S. Laimbacher

7. oHSV Genome Editing by Means of galK Recombineering
Laura Menotti, Valerio Leoni, Valentina Gatta , Biljana Petrovic, Andrea Vannini, Simona Pepe, Tatiana Gianni, and Gabriella Campadelli-Fiume

8. Rescue, Purification, and Characterization of a Recombinant HSV Expressing a Transgenic Protein
Andrea Vannini, Biljana Petrovic, Valentina Gatta, Valerio Leoni, Simona Pepe, Laura Menotti, Gabriella Campadelli-Fiume, and Tatiana Gianni

9. CRISPR/Cas9-Based Genome Editing of HSV
Thilaga Velusamy, Anjali Gowripalan, and David C. Tscharke

10. Latent/Quiescent Herpes Simplex Virus 1 Genome Detection by Fluorescent In Situ Hybridization (FISH)
Camille Cohen, Armelle Corpet, Mohamed Ali Maroui, Franceline Juillard, and Patrick Lomonte

11. Oligonucleotide Enrichment of HSV-1 Genomic DNA from Clinical Specimens for use in High-Throughput Sequencing
Mackenzie M. Shipley, Molly M. Rathbun, and Moriah L. Szpara

12. HSV Mutant Generation and Dual-Detection Methods for Gaining Insight into Latent/Lytic Cycles In Vivo
Nancy M. Sawtell and Richard L. Thompson

13. Phenotypic and Genotypic Testing of HSV-1 and HSV-2 Resistance to Antivirals
Andreas Sauerbrei and Kathrin Bohn-Wippert

14. Using Primary SCG Neuron Cultures to Study Molecular Determinants of HSV-1 Latency and Reactivation
Hui-Lan Hu, Kalanghad Puthankalam Srinivas, Ian Mohr, Tony T. Huang, and Angus C. Wilson

15. Characterization of Extracellular HSV-1 Virions by Proteomics
Roger Lippe

16. Analysis and Sorting of Individual HSV-1 Particles by Flow Virometry
Bita Khadivjam, Nabil El Bilali, and Roger Lippe

17. Isolation/Analysis of Extracellular Microvesicles from HSV-1-Infected Cells
Raquel Bello-Morales and Jose Antonio Lo pez-Guerrero

18. Conformational Change in Herpes Simplex Virus Entry Glycoproteins Detected by Dot Blot
Tri Komala Sari, Katrina A. Gianopulos, and Anthony V. Nicola

19. BioID Combined with Mass Spectrometry to Study Herpesvirus Protein-Protein Interaction Networks
Mujeeb R. Cheerathodi and David G. Meckes Jr.

20. Preparation of Herpes Simplex Virus Infected Primary Neurons for Transmission Electron Microscopy
Monica Miranda-Saksena, Ross Boadle, and Anthony L. Cunningham

21. Transmission Immuno-Electron Microscopy of Herpes Simplex Virus-1 Infected Dorsal Root Ganglia Neurons Sectioned in Growth Plane
Monica Miranda-Saksena, Ross Boadle, and Anthony L. Cunningham

22. Multi-Fluorescence Live Analysis of Herpes Simplex Virus Type-1 Replication
Michael Seyffert and Cornel Fraefel

23. Expression, Purification, and Crystallization of HSV-1 Glycoproteins for Structure Determination
Ellen M. White, Samuel D. Stampfer, and Ekaterina E. Heldwein

24. Expression, Purification, and Crystallization of Full-Length HSV-1 gB for Structure Determination
Rebecca S. Cooper and Ekaterina E. Heldwein

25. The Use of Microfluidic Neuronal Devices to Study the Anterograde Axonal Transport of Herpes Simplex Virus-1
Kevin Danastas, Anthony L. Cunningham, and Monica Miranda-Saksena

26. A Model of In Vivo HSV-1 DNA Transport using Murine Retinal Ganglion Cells
Jennifer H. LaVail

27. The Murine Intravaginal HSV-2 Challenge Model for Investigation of DNA Vaccines
Joshua O. Marshak, Lichun Dong, and David M. Koelle