Viral Vectors for Gene Therapy

This volume discusses protocols, ranging from vector production to delivery methods, used to execute gene therapy applications. Chapters are divided into four parts, and cover topics such as design, construction, and application of transcription activation-like effectors; multi-modal production of adeno-associated virus; construction of oncolytic herpes simplex virus; AAV-mediated gene delivery to the mouse liver; and intrathecal delivery of gene therapeutics by direct lumbar puncture in mice. 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.

Comprehensive and authoritative, Viral Vectors for Gene Therapy: Methods and Protocols is a valuable resource for researchers, clinicians, and students looking to utilize viral vectors in gene therapy experiments.

Basic Concepts in Viral Vector-Mediated Gene Therapy. - Design and Assembly of CRISPR/Cas9 Lentiviral and rAAV Vectors for Targeted Genome Editing. - Design, Construction, and Application of Transcription Activation-Like Effectors. - Practical Considerations for the use of DREADD and Other Chemogenetic Receptors to Regulate Neuronal Activity in the Mammalian Brain. - AAV Production using Baculovirus Expression Vector System. - Multimodal Production of Adeno-Associated Virus. - Generation of High Titer Pseudotyped Lentiviral Particles. - A Scalable Lentiviral Vector Production and Purification Method using Mustang Q Chromatography and Tangential Flow Filtration. - Current Use of Adenovirus Vectors and their Purification Methods. - Construction of Oncolytic Herpes Simplex Virus with Therapeutic Genes of Interest. - Poxviruses as Gene Therapy Vectors: Generating Poxviral Vectors Expressing Therapeutic Transgenes. - AAV Mediated Gene Delivery to the Mouse Liver. - Surgical Methods for Inner Ear Gene Delivery in Neonatal Mouse. - Gene Transfer to Mouse Kidney In Vivo. - Co-Delivery of a Short-Hairpin RNA and a shRNA-Resistant Replacement Gene with Adeno-Associated Virus: An Allele-Independent Strategy for Autosomal Dominant Retinal Disorders. - Localized Intra-Arterial Gene Delivery using AAV. - Stable Genetic Modification of Mesenchymal Stromal Cells Using Lentiviral Vectors. - Systemic Delivery of Adeno-Associated Viral Vectors in Mice and Dogs. - Intrathecal Delivery of AAV Vectors in Cynomolgus Macaques for CNS Gene Therapy and Gene Expression Analysis in Microdissected Motor Neurons. - Detailed Method for Intrathecal Delivery of Gene Therapeutics by Direct Lumbar Puncture in Mice. - Cerebromedullary Cistern Injection of Viral Vectors in Non-Human Primates.