University of Alberta Faculty of Medicine and Dentistry 3 articles published in JoVE Medicine Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts Rika Maruyama1, Aleksander Touznik1, Toshifumi Yokota1,2 1Department of Medical Genetics, University of Alberta Faculty of Medicine and Dentistry, 2Muscular Dystrophy Canada Research Chair, Department of Medical Genetics, University of Alberta Faculty of Medicine and Dentistry Various antisense oligonucleotides (AONs) have been shown to induce exon inclusion (splice modulation) and rescue SMN expression for spinal muscular atrophy (SMA). Here, we describe a protocol for AON lipotransfection to induce exon inclusion in the SMN2 gene and the evaluation methods to determine the efficacy in SMA patient fibroblasts. Immunology and Infection Cell Membrane Repair Assay Using a Two-photon Laser Microscope Joshua J. A. Lee1, Rika Maruyama1, Hidetoshi Sakurai2, Toshifumi Yokota1 1Department of Medical Genetics, University of Alberta Faculty of Medicine and Dentistry, 2Center for iPS Cell Research and Application, Kyoto University Cell membrane wounding via two-photon laser is a widely used method for assessing membrane resealing ability and can be applied to multiple cell types. Here, we describe a protocol for in vitro live-imaging of membrane resealing in dysferlinopathy patient cells following two-photon laser ablation. Medicine Multi-exon Skipping Using Cocktail Antisense Oligonucleotides in the Canine X-linked Muscular Dystrophy Bailey Miskew Nichols*1, Yoshitsugu Aoki*2, Mutsuki Kuraoka2, Joshua J.A. Lee1, Shin'ichi Takeda2, Toshifumi Yokota1 1Department of Medical Genetics, University of Alberta Faculty of Medicine and Dentistry, 2Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry Exon skipping is currently a most promising therapeutic option for Duchenne muscular dystrophy (DMD). To expand the applicability for DMD patients and to optimize the stability/function of the resulting truncated dystrophin proteins, a multi-exon skipping approach using cocktail antisense oligonucleotides was developed and we demonstrated systemic dystrophin rescue in a dog model.
