Main description:

The field of oligonucleotide therapeutics research is ripe with the prospect of new discoveries. In Therapeutic Oligonucleotides: Methods and Protocols, a selection of established and emerging methods for the application of oligonucleotides as therapeutics are presented, all providing the tools needed to inspire great changes in the field. Divided into twenty-one chapters, this detailed volume meticulously describes vital protocols for optimizing and improving cell uptake, such as photochemical internalization, modified cell penetrating peptides, antibody conjugates, and nanoparticles. Other chapters address quantitation of RNA therapeutics in cells, assaying gene knockdown, selecting the best target site and synthesis of various modified oligonucleotides. Written in the successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Therapeutic Oligonucleotides: Methods and Protocols serves as a timely resource for both professionals and novices pursuing research in this exciting and pioneering field.

Contents:

1. Therapeutic Oligonucleotides

John Goodchild

2. Dinucleotides Containing 3c-S-Phosphorothiolate Linkages

James W. Gaynor and Richard Cosstick

3. 2'-O,4'-C-Methyleneoxymethylene Bridged Nucleic Acids (2',4'-BNACOC)

Yoshiyuki Hari, Tetsuya Kodama, Takeshi Imanishi, and Satoshi Obika

4. A Non-covalent Peptide-based Strategy for Ex-vivo and In vivo Oligonucleotide Delivery

Laurence Crombez, May C. Morris, Frederic Heitz, and Gilles Divita

5. Cell Penetrating Peptides-based Strategies for the Delivery of Splice Redirecting Antisense Oligonucleotides

Samir El Andaloussi, Fatouma Said Hassane, Prisca Boisguerin, Rannar Sillard, Ulo Langel, and Bernard Lebleu

6. A Nanoparticle for Tumor Targeted Delivery of Oligomers

Xinrong Liu, Yi Wang, and Donald J. Hnatowich

7. Light-directed Delivery of Nucleic Acids

Sigurd Boe, Lina Prasmickaite, Birgit Engesaeter, and Eivind Hovig

8. Antibody Targeted siRNA Delivery

Masoud M. Toloue and Lance P. Ford

9. Aptamer-Drug Conjugation for Targeted Tumor Cell Therapy

Michael J. Donovan, Ling Meng, Tao Chen, Yunfei Zhang, Kwame Sefah, and Weihong Tan

10. Five-step Process for Screening Antisense Compounds for Efficacy: Gene Target IL-12Rb2

Nikki B. Marshall, Laura L. Hauck, and Dan V. Mourich

11. Diverse Small Non-coding RNAs in RNA Interference Pathways

Liande Li and Yi Liu

12. Quantification of siRNAs In vitro and In vivo

Angie Cheng, Alexander V. Vlassov, and Susan Magdaleno

13. Optimization of Transfection Conditions and Analysis of siRNA Potency Using Real-time PCR

Angie Cheng, Susan Magdaleno, and Alexander V. Vlassov

14. siRNA Knockdown of Gene Expression in Endothelial Cells

Emily Dennstedt and Brad Bryan

15. Using RNA Interference in Schistosoma mansoni

Rita Bhardwaj, Greice Krautz-Peterson, and Patrick J. Skelly

16. Performing the Labeled microRNA Pull-down (LAMP) Assay System: An Experimental Approach for High-throughput Identification of microRNA-target mRNAs

Ren-Jun Hsu and Huai-Jen Tsai

17. Synthesis, Purification, and Characterization of Oligoribonucleotides that Act as Agonists of TLR7 and/or TLR8

Tao Lan and Ekambar R. Kandimalla

18. Synthesis, Purification, and Characterization of Immune-Modulatory Oligodeoxynucleotides that Act as Agonists of Toll-like Receptor 9

Mallikarjuna Reddy Putta, Dong Yu, and Ekambar R. Kandimalla

19. Surface Plasmon Resonance Investigation of RNA Aptamer-RNA Ligand Interactions

Carmelo Di Primo, Eric Dausse, and Jean-Jacques Toulme

20. Practical Considerations for Analyzing Antigene RNAs (agRNAs): RNA Immunoprecipitation of Argonaute Protein

Jacob C. Schwartz and David R. Corey

21. Inhibition of Human Papillomavirus Expression Using DNAzymes

Maria Luisa Benitez-Hess, Pablo Reyes-Gutierrez, and Luis Marat Alvarez-Salas