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This volume provides a clear and detailed roadmap of how to design and execute a gene therapy experiment in order to obtain consistent results. Chapters in this book disseminate bits of unknown information that are important to consider during the course of experimentation and will answer questions such as: What delivery vehicle do you use?; How will you ensure that your vector retains stability?; What expression system best fits your needs?; What route will you choose to deliver your gene therapy agent?; How will you model the neurodegenerative disorder that you aim to investigate and what are the proven methods to treat these disorders in preclinical models? 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.
Authoritative and thorough, Gene Therapy for Neurological Disorders: Methods and Protocols, is a compilation of protocols and instructive chapters intended to give researchers, clinicians, and students of all levels, a foundation upon which future gene therapy experiments can be designed.
Introduction to Viral Vectors and Other Delivery Methods for Gene Therapy of the Nervous System.- Delivering Transgenic DNA Exceeding the Carrying Capacity of AAV Vectors.- Expression of Multiple Functional RNAs or Proteins from One Viral Vector.- Regulated Gene Therapy.- Design of shRNA and miRNA for Delivery to the CNS.- Tissue Specific Promoters in the CNS.- Small Scale Recombinant Adeno-Associated Virus Purification.- Lentivirus Production and Purification.- Viral Vector Production: Adenovirus.- Controlling AAV Tropism in the Nervous System with Natural and Engineered Capsids.- Altering Tropism of rAAV by Directed Evolution.- Altering Entry Site Preference of Lentiviral Vectors into Neuronal Cells by Pseudotyping with Envelope Glycoproteins.- Directed Evolution of Adenoviruses.- Intraparenchymal Stereotaxic Delivery of rAAV and Special Considerations in Vector Handling.- MRI-Guided Delivery of Viral Vectors.- Systemic Gene Therapy for Targeting the CNS.- Widespread Neuronal Transduction of the Rodent CNS via Neonatal Viral Injection.- AAV-Mediated Gene Transfer to Dorsal Root Ganglion.- Gene Therapy of the Peripheral Nervous System: The Enteric Nervous System.- Gene Therapy of the Peripheral Nervous System: Celiac Ganglia.- Convection Enhanced Delivery of Recombinant Adeno-Associated Virus into the Mouse Brain.- Non-Viral Gene Therapy of the Nervous System: Electroporation.- Non-Viral, Lipid-Mediated DNA and mRNA Gene Therapy of the Central Nervous System (CNS): Chemical-Based Transfection.- Ex Vivo Gene Therapy Using Human Mesenchymal Stem Cells to Deliver Growth Factors in the Skeletal Muscle of a Familial ALS Rat Model.- Gene Therapy Models of Alzheimer's Disease and Other Dementias.- Viral Vector-Based Modeling of Neurodegenerative Disorders: Parkinson's Disease.- Gene Therapy-Based Modeling of Neurodegenerative Disorders: Huntington's Disease.- Gene Therapy for the Treatment of Neurological Disorders: AmyotrophicLateral Sclerosis.- Stereotaxic Surgical Targeting of the Non-Human Primate Causate and Putamen: Gene Therapy for Huntington's Disease.- Gene-Therapy for the Treatment of Neurological Disorders: Metabolic Disorders.- Gene Therapy for the Treatment of Neurological Disorders: Central Nervous System Neoplasms.- AAV2-Neurturin for Parkinson's Disease: What Lessons Have We Learned?.
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