Gapmers Methods and Protocols

This volume presents a comprehensive collection of detailed state-of-the-art protocols for gapmer-mediated RNA knockdown from leaders in the field. Chapters detail a historical and contemporary perspective of RNase H-inducing antisense oligonucleotides called gapmers and gapmer-mediated RNA knockdown for basic research and therapies. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

 Authoritative and cutting-edge, Gapmers: Methods and Protocols aims to provide diverse applications of gapmers along with protocols that will assist readers in moving the frontier.

Invention and Early History.- Development and Clinical Applications of Antisense Oligonucleotide Gapmers.- Knocking Down Long Non-coding RNAs using Antisense Oligonucleotide Gapmers.- Development of Antisense Oligonucleotide Gapmers for the Treatment of Huntington’s Disease.- Development of Antisense Oligonucleotide Gapmers for the Treatment of Dyslipidemia and Lipodystrophy.- Inotersen for the Treatment of Hereditary Transthyretin Amyloidosis.- Degradation of Toxic RNA in Myotonic Dystrophy Using Gapmer Antisense Oligonucleotides.- DNA/RNA Heteroduplex Oligonucleotide for Highly Efficient Gene Silencing.- Tips for Successful lncRNA Knockdown using Gapmers.- Calcium-Mediated In Vitro Transfection Technique of Oligonucleotides with Broad Chemical Modification Compatibility.- Evaluating the Knockdown Activity of MALAT1 ENA Gapmers in vitro.- Albumin-binding Fatty Acid-modified Gapmer Antisense Oligonucleotides for Modulation of Pharmacokinetics.- The use of Gapmers for in vivo Suppression of Hepatic mRNA Targets.- Development of LNA Gapmer Oligonucleotide-Based Therapy for ALS/ FTD Caused by the C9orf72 Repeat Expansion.- Targeted Gene Silencing in Malignant Hematolymphoid Cells using GapmeR.- Gapmer Antisense Oligonucleotides to Selectively Suppress the Mutant allele in COL6A Genes in Sominant Ullrich Congenital Muscular Dystrophy.- Detection of Locked Nucleic Acid Gapmers from Mouse Samples using ELISA.