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. 1. Therapeutic Oligonucleotides John Goodchild 2. Dinucleotides Containing 3-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 Be, Lina Prasmickaite, Birgit Engester, 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 Toulm 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 Mara Luisa Bentez-Hess, Pablo Reyes-Gutirrez, and Luis Marat Alvarez-Salas
