Retinal Gene Therapy (Methods in Molecular Biology) de Camiel J.F. Boon

Retinal Gene Therapy: Methods and Protocols: Methods in Molecular Biology, cartea 1715

Camiel J.F. Boon

Jan Wijnholds

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.

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Cuprins

Small Scale Production of Recombinant Adeno-Associated Viral Vectors for Gene Delivery to the Nervous System.- Small and Micro-Scale Recombinant Adeno-Associated Virus Production and Purification for Ocular Gene Therapy Applications.- Design and Development of AAV-Based Gene Supplementation Therapies for Achromatopsia and Retinitis Pigmentosa.- Development of Multigenic Lentiviral Vectors for Cell-Specific Expression of Antiangiogenic miRNAs and Protein Factors.- Design and In Vitro Use of Antisense Oligonucleotides to Correct Pre-mRNA Splicing Defects in Inherited Retinal Dystrophies.- Three-Dimensional Co-Culture Bioassay for Screening of Retinal Gene Delivery Systems.- Retinal Gene Therapy for Choroideremia: In Vitro Testing for Gene Augmentation Using an Adeno-Associated Viral (AAV) Vector.- In Vivo Electroporation of Developing Mouse Retina.- Methods for In Vivo CRISPR/Cas Editing of the Adult Murine Retina.- AAV Gene Augmentation Therapy for CRB1-Associated Retinitis Pigmentosa.- Dual AAV Vectors for Stargardt Disease.- Optogenetic Retinal Gene Therapy with the Light Gated GPCR Vertebrate Rhodopsin.- CRISPR Repair Reveals Causative Mutation in a Preclinical Model of Retinitis Pigmentosa: A Brief Methodology.- In-Depth Functional Analysis of Rodents by Full-Field Electroretinography.- Advanced Ocular Injection Techniques for Therapy Approaches.- Neutralizing Antibodies Against Adeno-Associated Virus (AAV): Measurement and Influence on Retinal Gene Delivery.- Screening for Neutralizing Antibodies against Natural and Engineered AAV Capsids in Non-Human Primate Retinas.- Subretinal and Intravitreal Retinal Injections in Monkeys.- Production of iPS-derived Human Retinal Organoids for Use in Transgene Expression Assays.- AAV Serotype Testing on Cultured Human Donor Retinal Explants.- Human Retinal Explant Culture for Ex-Vivo Validation of AAV Gene Therapy.- Visual Acuity Testing Before and After Intravitreal Injection of rAAV2- ND4 in Patients.- Recordingand Analysis of the Human Clinical Electroretinogram.- Recording and Analysis of Goldmann Kinetic Visual Fields.- Measuring Central Retinal Sensitivity Using Microperimetry.- Inspection of the Human Retina by Optical Coherence Tomography.- Vector Shedding and Immunogenicity Sampling for Retinal Gene Therapy.

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Retinal Gene Therapy: Methods and Protocols: Methods in Molecular Biology, cartea 1715

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