Exon Skipping and Inclusion Therapies

Part I: Basics and Introductions

1. Invention and Early History of Exon Skipping and Splice Modulation

Kenji Rowel Q. Lim and Toshifumi Yokota

2. An Overview of Recent Advances and Clinical Applications of Exon Skipping and Splice Modulation for Muscular Dystrophy and Various Genetic Diseases

Merryl Rodrigues and Toshifumi Yokota

3. Recent Advances and Clinical Applications of Exon Inclusion for Spinal Muscular Atrophy

4. Nusinersen in the Treatment of Spinal Muscular Atrophy

Kara Goodkey, Tejal Aslesh, Rika Maruyama, and Toshifumi Yokota

Part II: Design of AONs and Strategy

5. Tips to Design Effective Splice-switching Antisense Oligonucleotides for Exon Skipping and Exon Inclusion

6. Antisense Oligonucleotide Targeting of 3'UTR of mRNA for Expression Knockdown

Golnoush Golshirazi, Lukasz Ciszewski, Ngoc Lu-Nguyen, and Linda Popplewell

Part III: In Vitro Evaluation of Exon Skipping for DMD

7. Quantitative Evaluation of Exon Skipping in Immortalized Muscle Cells In Vitro

Kenji Rowel Q. Lim and Toshifumi Yokota

8. Direct Reprogramming of Human DMD Fibroblasts into Myotubes for In Vitro Evaluation of Antisense-m Exon Skipping and Exons 45-55 Skipping Accompanied by Rescue of Dystrophin Expression

Joshua JA Lee, Takashi Saito, William Duddy, Shin'ichi Takeda, and Toshifumi Yokota

9. In vitro Multi-exon Skipping by Antisense PMOs in Dystrophic Dog and Exon 7-Deleted DMD Patient

Akinori Nakamura, Yoshitsugu Aoki, Maria Tsoumpra, Toshifumi Yokota, and Shin'ichi Takeda

10. Creation of DMD Muscle Cell Model using CRISPR-Cas9 Genome Editing to Test the Efficacy of Antisense-mediated Exon Skipping

Rika Maruyama and Toshifumi Yokota

11. In vitro Evaluation of Exon Skipping in Disease Specific iPSC-derived Myocytes

Mingming Zhao, Emi Shoji, and Hidetoshi Sakurai

12. 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, and Akitsu Hotta

13. Skipping of Duplicated Dystrophin Exons: in vitro Induction and Assessment

Kane Greer, Sue Fletcher, and Steve D Wilton

Part IV: In Vivo Evaluation of Exon Skipping for DMD

14. In Vivo Evaluation of Dystrophin Exon Skipping in mdx Mice

Bo Wu, Mingxing Wang, Sapana Shah, and Qi Long Lu

15. Exon 51 Skipping Quantification by Digital Droplet PCR in del52hDMD/mdx Mice

Monika Hiller, Pietro Spitali, Nicole Datson, and Annemieke Aartsma-Rus

16. Systemic Injection of Peptide-PMOs into Humanized DMD Mice and Detection by RT-PCR and ELISA

Dyanna Melo, Rika Maruyama, and Toshifumi Yokota

17. In vivo Evaluation of Single- and Multi-exon Skipping in mdx52 Mice

Yoshitaka Mizobe, Shouta Miyatake, Hotake Takizawa, Yuko Hara, Toshifumi Yokota, Akinori Nakamura, Shin'ichi Takeda, and Yoshitsugu Aoki

18. A Novel Zebrafish Model for Assessing In Vivo Delivery of Morpholino Oligomers

Joseph Kim, Karl Clark, Carrie Barton, Robert Tanguay, and Hong Moulton

Part V: Enhancing the Efficacy

19. Validation and Detection of Exon Skipping Boosters in DMD Patient Cell Models and mdx Mouse

Barthelemy. Florian, Wang. Dereck, Nelson. Stanley F., Miceli, M. Carr