Antisense RNA Design, Delivery, and Analysis

Part I: Overview

1. Introduction and History of the Chemistry of Nucleic Acids Therapeutics

Michael J. Gait and Sudhir Agrawal

2. Antisense RNA Therapeutics: A Brief Overview

Virginia Arechavala-Gomeza and Alejandro Garanto

Part II: Design and Formulation of Antisense Technology

3. Design of Bifunctional Antisense Oligonucleotides for Exon Inclusion

Haiyan Zhou

4. Design and Delivery of SINEUP: A New Modular Tool to Increase Protein Translation

Michele Arnoldi, Giulia Zarantonello, Stefano Espinoza, Stefano Gustincich, Francesca Di Leva, and Marta Biagioli

5. How to Design U1 snRNA Molecules for Splicing Rescue

Liliana Matos, Juliana I. Santos, Ma Francisca Coutinho, and Sandra Alves

6. Conjugation of Nucleic Acids and Drugs to Gold Nanoparticles

Paula Miln-Rois, Ciro Rodriguez-Diaz, Milagros Castellanos, and lvaro Somoza

7. Determination of Optimum Ratio of Cationic Polymers and Small Interfering RNA with Agarose Gel Retardation Assay

Omer Aydin, Dilek Kanarya, Ummugulsum Yilmaz, and Cansu Umran Tunc

8. Generation of Protein-Phosphorodiamidate Morpholino Oligomer Conjugates for Efficient Cellular Delivery via Anthrax Protective Antigen

Valentina Palacio-Castaeda, Roland Brock, and Wouter P.R. Verdurmen

Part III: In Vitro Model Systems

9. Development and Use of Cellular Systems to Assess and Correct Splicing Defects

Nuria Surez-Herrera, Tomasz Z. Tomkiewicz, Alejandro Garanto, and Rob W.J. Collin

10. Modeling Splicing Variants Amenable to Antisense Therapy by Use of CRISPR-Cas9-Based Gene Editing in HepG2 Cells

Arstides Lpez-Mrquez, Ainhoa Martnez-Pizarro, Beln Prez, Eva Richard, and Lourdes R. Desviat

11. In Vitro Models for the Evaluation of Antisense Oligonucleotides in Skin

Jeroen Bremer and Peter C. van den Akker

12. In Vitro Delivery of PMOs in Myoblasts by Electroporation

Remko Goossens and Annemieke Aartsma-Rus

13. Rapid Determination of MBNL1 Protein Levels by Quantitative Dot Blot for Evaluation of Antisense Oligonucleotides in Myotonic Dystrophy Myoblasts

Nerea Moreno-Cervera, Irene Gonzalez-Martinez, Rubn Artero, and Estefana Cerro-Herreros

14. Evaluation of Exon Skipping and Dystrophin Restoration in In Vitro Models of Duchenne Muscular Dystrophy

Andrea Lpez-Martnez, Patricia Soblechero-Martn, and Virginia Arechavala-Gomeza

15. Generation of Human iPSC-Derived Myotubes to Investigate RNA-Based