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 Yokota^{1,2 1}Department 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 Yokota^{1 1}Department 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 Yokota^{1 1}Department 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.