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University of Alberta Faculty of Medicine and Dentistry

3 ARTICLES PUBLISHED IN JoVE

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.

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

Evaluation of Exon Inclusion Induced by Splice Switching Antisense Oligonucleotides in SMA Patient Fibroblasts

Rika Maruyama¹, Aleksander Touznik¹, Toshifumi Yokota^1,2

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.