Full Description
This volume covers the latest advancements in the study of retinal gene therapy and addresses critical aspects of the gene therapy development pipeline. The chapters in this book are organized into four parts. Part One focuses on vector development and design, and talks about the latest approaches used to enhance the specificity and efficiency of gene delivery to retinal cells. Part Two presents a series of in vitro assays that are important for validating retinal gene therapies. Part Three discusses in vivo proof-of-concept studies and therapy validation in animal models. Part Four addresses clinical validation and presents methods for assessing visual function in human subjects undergoing retinal gene therapy. 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, Retinal Gene Therapy: Methods and Protocols is a valuable resource for researchers, clinicians, and students who are interested in learning more about this rapidly evolving field and looking for new research directions and collaborations across disciplines.
Contents
Selecting Novel Retino-Tropic AAV Capsids by Directed Evolution.- Evaluation of CRISPR/CasRx-Mediated VEGF mRNA Knockdown in Mouse Retina.- Optimized Retinal Cell Dissociation for Downstream Analysis of Cone Photoreceptors.- Development of In Vitro Potency Assays for Gene Therapy Products Targeting cGMP-Related Retinopathies.- Quantification of Drusen-Like Deposits in Human Pluripotent Stem Cell-Derived Retinal Pigment Epithelium Cells Using Confocal Microscopy.- Ex Vivo Culture and Formalin-Fixed Paraffin Embedding of Murine Retinal Explants.- Culture of Human Retinal Explants as an Ex Vivo Model for Retinal Gene Therapy.- Electrophysiological Characterization of Photoreceptor-Like Cells in Retinal Organoids Using Whole-Cell Patch Clamp Method.- Generation and Characterization of Patient-Derived Human Pluripotent Stem Cell Lines for Retinal Gene Therapy.- Transduction of AAV Vectors in iPSC-Derived Retinal Organoids.- Intravitreal Injections in Mouse Models of Eye Disease.- Behavioral Assessment of Vision in Dogs Using a Four-Alternative Choice-Based Testing Device.- Behavioral Vision Testing Using an Automated Observational Tracking System (VisioBox) to Obtain a Measure of Visual Acuity in Larval Zebrafish.- Design and Validation of Mirtron-Mediated RNA Interference for Gene Therapy.- Applications of Fundus-Tracked Perimetry in the Assessment of Visual Sensitivity.- Assessing Visual Function in Retinal Gene Therapy.
