This volume explores the latest advancements in the field of RNA therapeutics, with a particular focus on the development of exon skipping and inclusion therapies as targeted treatments for genetic diseases, as well as the evaluation methods used post-treatment. The chapters in this book cover both established methods and novel techniques used my researchers working on developing therapies that have proven to be promising for several diseases, including Duchenne muscular dystrophy (DMD), spinal muscular atrophy (SMA), and other genetic disorders characterized by abnormal splicing events. 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 thorough, Splice Switching: Methods and Protocols is a valuable resource for researchers across various fields. This book will help in the advancement of basic RNA biology research and aid in accelerating therapeutic developments.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

DG9-Conjugated Morpholino-Based Exon 51 Skipping Therapy for Duchenne Muscular Dystrophy.- DG9-Conjugated Morpholino Rescues Phenotype in Spinal Muscular Atrophy Mice.- Cardio-Physiological Evaluations in A Dog Model with Duchenne Muscular Dystrophy.- Splice Modulation Studies Using Urine-Derived Cells.- Lipidomic Sample Preparation to Detect Phosphatidylcholine Alterations in Skeletal Muscle.- Evaluation for the Efficacy of Exon Skipping Therapy Using In Vivo Assessment of Skeletal Muscle Function and Fragility.- Characterization and Phenotypic Assessment of Zebrafish Disease Models for Splice Modulation Studies.- Preparation of Size-Defined PEG-Grafted Copolymers as a Polymeric Nanoruler for Size Optimization in Passive Targeting.- Splice Modulation Studies Using Reporter Mice.- Clinical Variant Databases and Machine Learning Prediction Supporting Genomic Medicine.- In Vivo Evaluation of CTG Repeat-Affected Muscle Pathology in a Myotonic Dystrophy Model Mouse Using Electromyography and Fluorescence In Situ Hybridization.- Utilization of Induced Pluripotent Stem Cell-Derived Neurons to Investigate Splice-Modification Efficacy of Splice-Switching Drug Candidates.- Development of Electric Field Stimulation (EFS)-Mediated Skeletal Muscle Training with iPSCs.- Neurological Behavioral Assessment of Exon Skipping Therapy for Neuromuscular Disorder Using Displaced Object Recognition Test, Open Field Test, and Three-Chamber Social Approach Test.- Analysis of Prefrontal-Amygdala Synaptic Functions Using Optogenetics and Path-Clamp Recording.- Digital Outcome Measures for Gait and Spontaneous Locomotor Activity in Canine Dystrophic Model.