Therapeutic Oligonucleotides

This volume in the Methods in Molecular Biology(TM) series presents established and emerging methods for the application of oligonucleotides as therapeutics. Includes protocols for optimizing cell uptake, assaying gene knockdown, target site selection and more.

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™ 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.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras

Inhalt

Therapeutic Oligonucleotides.- Dinucleotides Containing 3¢-S-Phosphorothiolate Linkages.- 2'-O,4'-C-Methyleneoxymethylene Bridged Nucleic Acids (2',4'-BNACOC).- A Non-covalent Peptide-based Strategy for Ex-vivo and In vivo Oligonucleotide Delivery.- Cell Penetrating Peptides-based Strategies for the Delivery of Splice Redirecting Antisense Oligonucleotides.- A Nanoparticle for Tumor Targeted Delivery of Oligomers.- Light-directed Delivery of Nucleic Acids.- Antibody Targeted siRNA Delivery.- Aptamer-Drug Conjugation for Targeted Tumor Cell Therapy.- Five-step Process for Screening Antisense Compounds for Efficacy: Gene Target IL-12Rb2.- Diverse Small Non-coding RNAs in RNA Interference Pathways.- Quantification of siRNAs In vitro and In vivo.- Optimization of Transfection Conditions and Analysis of siRNA Potency Using Real-time PCR.- siRNA Knockdown of Gene Expression in Endothelial Cells.- Using RNA Interference in Schistosoma mansoni.- Performing the Labeled microRNA Pull-down (LAMP) Assay System: An Experimental Approach for High-throughput Identification of microRNA-target mRNAs.- Synthesis, Purification, and Characterization of Oligoribonucleotides that Act as Agonists of TLR7 and/or TLR8.- Synthesis, Purification, and Characterization of Immune-Modulatory Oligodeoxynucleotides that Act as Agonists of Toll-like Receptor 9.- Surface Plasmon Resonance Investigation of RNA Aptamer-RNA Ligand Interactions.- Practical Considerations for Analyzing Antigene RNAs (agRNAs): RNA Immunoprecipitation of Argonaute Protein.- Inhibition of Human Papillomavirus Expression Using DNAzymes