Exon Skipping and Inclusion Therapies: Methods and Protocols

Front Matter ....Pages i-xv
Front Matter ....Pages 1-1
 Invention and Early History of Exon Skipping and Splice Modulation  (Kenji Rowel Q. Lim, Toshifumi Yokota)....Pages 3-30
 An Overview of Recent Advances and Clinical Applications of Exon Skipping and Splice Modulation for Muscular Dystrophy and Various Genetic Diseases  (Merryl Rodrigues, Toshifumi Yokota)....Pages 31-55
 Recent Advances and Clinical Applications of Exon Inclusion for Spinal Muscular Atrophy  (Hae-Won Son, Toshifumi Yokota)....Pages 57-68
 Nusinersen in the Treatment of Spinal Muscular Atrophy  (Kara Goodkey, Tejal Aslesh, Rika Maruyama, Toshifumi Yokota)....Pages 69-76
Front Matter ....Pages 77-77
 Tips to Design Effective Splice-Switching Antisense Oligonucleotides for Exon Skipping and Exon Inclusion  (Rika Maruyama, Toshifumi Yokota)....Pages 79-90
 Antisense Oligonucleotide Targeting of 3’-UTR of mRNA for Expression Knockdown  (Golnoush Golshirazi, Lukasz Ciszewski, Ngoc Lu-Nguyen, Linda Popplewell)....Pages 91-124
Front Matter ....Pages 125-125
 Quantitative Evaluation of Exon Skipping in Immortalized Muscle Cells In Vitro  (Kenji Rowel Q. Lim, Toshifumi Yokota)....Pages 127-139
 Direct Reprogramming of Human DMD Fibroblasts into Myotubes for In Vitro Evaluation of Antisense-Mediated Exon Skipping and Exons 45–55 Skipping Accompanied by Rescue of Dystrophin Expression  (Joshua J. A. Lee, Takashi Saito, William Duddy, Shin’ichi Takeda, Toshifumi Yokota)....Pages 141-150
 In Vitro Multiexon Skipping by Antisense PMOs in Dystrophic Dog and Exon 7-Deleted DMD Patient  (Akinori Nakamura, Yoshitsugu Aoki, Maria Tsoumpra, Toshifumi Yokota, Shin’ichi Takeda)....Pages 151-163
 Creation of DMD Muscle Cell Model Using CRISPR-Cas9 Genome Editing to Test the Efficacy of Antisense-Mediated Exon Skipping  (Rika Maruyama, Toshifumi Yokota)....Pages 165-171
 In Vitro Evaluation of Exon Skipping in Disease-Specific iPSC-Derived Myocytes  (Mingming Zhao, Emi Shoji, Hidetoshi Sakurai)....Pages 173-189
 Restoration of Dystrophin Protein Expression by Exon Skipping Utilizing CRISPR-Cas9 in Myoblasts Derived from DMD Patient iPS Cells  (Masataka Ifuku, Kumiko A. Iwabuchi, Masami Tanaka, Mandy Siu Yu Lung, Akitsu Hotta)....Pages 191-217
 Skipping of Duplicated Dystrophin Exons: In Vitro Induction and Assessment  (Kane Greer, Sue Fletcher, Steve D. Wilton)....Pages 219-228
Front Matter ....Pages 229-229
 In Vivo Evaluation of Dystrophin Exon Skipping in mdx Mice  (Bo Wu, Mingxing Wang, Sapana Shah, Qi Long Lu)....Pages 231-247
 Exon 51 Skipping Quantification by Digital Droplet PCR in del52hDMD/mdx Mice  (Monika Hiller, Pietro Spitali, Nicole Datson, Annemieke Aartsma-Rus)....Pages 249-262
 Systemic Injection of Peptide-PMOs into Humanized DMD Mice and Evaluation by RT-PCR and ELISA  (Dyanna Melo, Rika Maruyama, Toshifumi Yokota)....Pages 263-273
 In Vivo Evaluation of Single-Exon and Multiexon Skipping in mdx52 Mice  (Yoshitaka Mizobe, Shouta Miyatake, Hotake Takizawa, Yuko Hara, Toshifumi Yokota, Akinori Nakamura et al.)....Pages 275-292
 A Novel Zebrafish Model for Assessing In Vivo Delivery of Morpholino Oligomers  (Joseph Kim, Karl Clark, Carrie Barton, Robert Tanguay, Hong Moulton)....Pages 293-306
Front Matter ....Pages 307-307
 Validation and Detection of Exon Skipping Boosters in DMD Patient Cell Models and mdx Mouse  (Florian Barthelemy, Dereck Wang, Stanley F. Nelson, M. Carrie Miceli)....Pages 309-326
 Use of Glucose–Fructose to Enhance the Exon Skipping Efficacy  (Gang Han, Xianjun Gao, Haifang Yin)....Pages 327-342
 Systemic Intravenous Administration of Antisense Therapeutics for Combinatorial Dystrophin and Myostatin Exon Splice Modulation  (Ngoc Lu-Nguyen, George Dickson, Alberto Malerba)....Pages 343-354
 The Assembly of Fluorescently Labeled Peptide–Oligonucleotide Conjugates via Orthogonal Ligation Strategies  (John Karas, Bradley J. Turner, Fazel Shabanpoor)....Pages 355-363
 In Vivo Evaluation of Multiple Exon Skipping with Peptide-PMOs in Cardiac and Skeletal Muscles in Dystrophic Dogs  (Rika Maruyama, Yoshitsugu Aoki, Shin’ichi Takeda, Toshifumi Yokota)....Pages 365-379
 Use of Tricyclo-DNA Antisense Oligonucleotides for Exon Skipping  (Karima Relizani, Aurelie Goyenvalle)....Pages 381-394
 Optimization of 2′,4′-BNA/LNA-Based Oligonucleotides for Splicing Modulation In Vitro  (Takenori Shimo, Satoshi Obika)....Pages 395-411
Front Matter ....Pages 413-413
 Pre-mRNA Splicing Modulation by Antisense Oligonucleotides  (Natalia N. Singh, Diou Luo, Ravindra N. Singh)....Pages 415-437
 In Vitro Evaluation of Antisense-Mediated Exon Inclusion for Spinal Muscular Atrophy  (Aleksander Touznik, Rika Maruyama, Toshifumi Yokota)....Pages 439-454
 Systemic and ICV Injections of Antisense Oligos into SMA Mice and Evaluation  (Tejal Aslesh, Rika Maruyama, Toshifumi Yokota)....Pages 455-465
 Morpholino-Mediated Exon Inclusion for SMA  (Haiyan Zhou, Francesco Muntoni)....Pages 467-477
Front Matter ....Pages 479-479
 Exon Skipping by Ultrasound-Enhanced Delivery of Morpholino with Bubble Liposomes for Myotonic Dystrophy Model Mice  (Yoichi Negishi, Yoko Endo-Takahashi, Shoichi Ishiura)....Pages 481-487
 Dysferlin Exon 32 Skipping in Patient Cells  (Florian Barthélémy, Sébastien Courrier, Nicolas Lévy, Martin Krahn, Marc Bartoli)....Pages 489-496
 Morpholino-Mediated Exon Skipping Targeting Human ACVR1/ALK2 for Fibrodysplasia Ossificans Progressiva  (Rika Maruyama, Toshifumi Yokota)....Pages 497-502
 Exon Skipping of FcεRIβ for Allergic Diseases  (Greer K. Arthur, Glenn Cruse)....Pages 503-518
 Antisense Oligonucleotide Design and Evaluation of Splice-Modulating Properties Using Cell-Based Assays  (Ralph Slijkerman, Hannie Kremer, Erwin van Wijk)....Pages 519-530
 Antisense-Mediated Splice Modulation to Reframe Transcripts  (Matthias Titeux, Sandrina Turczynski, Nathalie Pironon, Alain Hovnanian)....Pages 531-552
 Exon Skipping Using Antisense Oligonucleotides for Laminin-Alpha2-Deficient Muscular Dystrophy  (Yuko Hara, Yoshitaka Mizobe, Shouta Miyatake, Hotake Takizawa, Tetsuya Nagata, Toshifumi Yokota et al.)....Pages 553-564
 Correction to: Antisense-Mediated Splice Modulation to Reframe Transcripts  (Matthias Titeux, Sandrina Turczynski, Nathalie Pironon, Alain Hovnanian)....Pages E1-E1
Back Matter ....Pages 565-569