Retinal Gene Therapy: Methods and Protocols

Description

This detailed volume describes a spectrum of methods and protocols that can be used for the bench-to-bedside development and evaluation of retinal gene therapy. Methods for the successful delivery of these gene therapy vector systems to the retina are examined, as well as assays to test the efficacy in vitro in cell cultures, for gene augmentation and gene editing in vivo on rodents, pigs, and monkey retinas, and on human retinal explants as well as in human clinical studies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Retinal Gene Therapy: Methods and Protocols provides a wide range of readers from students to research experts with vital information on ocular gene therapy vector technology, in vitro and in vivo biological assays, and clinical protocols, to promote further studies for the benefit of children and adults with inherited retinal disease. Front Matter ....Pages i-xvi Front Matter ....Pages 1-1 Small Scale Production of Recombinant Adeno-Associated Viral Vectors for Gene Delivery to the Nervous System (Joost Verhaagen, Barbara Hobo, Erich M. E. Ehlert, Ruben Eggers, Joanna A. Korecka, Stefan A. Hoyng et al.)....Pages 3-17 Small and Micro-Scale Recombinant Adeno-Associated Virus Production and Purification for Ocular Gene Therapy Applications (Christopher A. Reid, Daniel M. Lipinski)....Pages 19-31 Design and Development of AAV-based Gene Supplementation Therapies for Achromatopsia and Retinitis Pigmentosa (Christian Schön, Elvir Becirovic, Martin Biel, Stylianos Michalakis)....Pages 33-46 Development of Multigenic Lentiviral Vectors for Cell-Specific Expression of Antiangiogenic miRNAs and Protein Factors (Anne Louise Askou, Thomas J. Corydon)....Pages 47-60 Design and In Vitro Use of Antisense Oligonucleotides to Correct Pre-mRNA Splicing Defects in Inherited Retinal Dystrophies (Alejandro Garanto, Rob W. J. Collin)....Pages 61-78 Three-Dimensional Co-Culture Bioassay for Screening of Retinal Gene Delivery Systems (Ding Wen Chen, Kathleen Pauloff, Marianna Foldvari)....Pages 79-88 Retinal Gene Therapy for Choroideremia: In Vitro Testing for Gene Augmentation Using an Adeno-Associated Viral (AAV) Vector (Maria I. Patrício, Robert E. MacLaren)....Pages 89-97 Front Matter ....Pages 99-99 In Vivo Electroporation of Developing Mouse Retina (Jimmy de Melo, Seth Blackshaw)....Pages 101-111 Methods for In Vivo CRISPR/Cas Editing of the Adult Murine Retina (Sandy S. Hung, Fan Li, Jiang-Hui Wang, Anna E. King, Bang V. Bui, Guei-Sheung Liu et al.)....Pages 113-133 AAV Gene Augmentation Therapy for CRB1-Associated Retinitis Pigmentosa (C. Henrique Alves, Jan Wijnholds)....Pages 135-151 Dual AAV Vectors for Stargardt Disease (Ivana Trapani)....Pages 153-175 Optogenetic Retinal Gene Therapy with the Light Gated GPCR Vertebrate Rhodopsin (Benjamin M. Gaub, Michael H. Berry, Meike Visel, Amy Holt, Ehud Y. Isacoff, John G. Flannery)....Pages 177-189 CRISPR Repair Reveals Causative Mutation in a Preclinical Model of Retinitis Pigmentosa: A Brief Methodology (Wen-Hsuan Wu, Yi-Ting Tsai, Sally Justus, Galaxy Y. Cho, Jesse D. Sengillo, Yu Xu et al.)....Pages 191-205 In-Depth Functional Analysis of Rodents by Full-Field Electroretinography (Vithiyanjali Sothilingam, Regine Mühlfriedel, Naoyuki Tanimoto, Mathias W. Seeliger)....Pages 207-213 Advanced Ocular Injection Techniques for Therapy Approaches (Regine Mühlfriedel, Marina Garcia Garrido, Christine Wallrapp, Mathias W. Seeliger)....Pages 215-223 Neutralizing Antibodies Against Adeno-Associated Virus (AAV): Measurement and Influence on Retinal Gene Delivery (Mélissa Desrosiers, Deniz Dalkara)....Pages 225-238 Screening for Neutralizing Antibodies Against Natural and Engineered AAV Capsids in Nonhuman Primate Retinas (Timothy P. Day, Leah C. Byrne, John G. Flannery, David V. Schaffer)....Pages 239-249 Subretinal and Intravitreal Retinal Injections in Monkeys (Daniyar Dauletbekov, K. Ulrich Bartz-Schmidt, M. Dominik Fischer)....Pages 251-257 Front Matter ....Pages 259-259 Production of iPS-Derived Human Retinal Organoids for Use in Transgene Expression Assays (Peter M. Quinn, Thilo M. Buck, Charlotte Ohonin, Harald M. M. Mikkers, Jan Wijnholds)....Pages 261-273 AAV Serotype Testing on Cultured Human Donor Retinal Explants (Thilo M. Buck, Lucie P. Pellissier, Rogier M. Vos, Elon H. C. van Dijk, Camiel J. F. Boon, Jan Wijnholds)....Pages 275-288 Human Retinal Explant Culture for Ex Vivo Validation of AAV Gene Therapy (Harry O. Orlans, Thomas L. Edwards, Samantha R. De Silva, Maria I. Patrício, Robert E. MacLaren)....Pages 289-303 Visual Acuity Testing Before and After Intravitreal Injection of rAAV2-ND4 in Patients (Bin Li, Chenmian Wu)....Pages 305-311 Recording and Analysis of the Human Clinical Electroretinogram (Mathieu Gauvin, Allison L. Dorfman, Pierre Lachapelle)....Pages 313-325 Recording and Analysis of Goldmann Kinetic Visual Fields (Mays Talib, Gislin Dagnelie, Camiel J. F. Boon)....Pages 327-338 Measuring Central Retinal Sensitivity Using Microperimetry (Mays Talib, Jasleen K. Jolly, Camiel J. F. Boon)....Pages 339-349 Inspection of the Human Retina by Optical Coherence Tomography (Thomas Theelen, Michel M. Teussink)....Pages 351-358 Vector Shedding and Immunogenicity Sampling for Retinal Gene Therapy (Alun R. Barnard, Anna N. Rudenko, Robert E. MacLaren)....Pages 359-371 Back Matter ....Pages 373-380