Extracellular RNA: Methods and Protocols

Description

This volume provides a collection of methods and protocols detailing extracellular RNA and hopes to be a useful resource for those interested in this exciting and emerging field. Chapters are divided into four sections covering an overview of the field, approaches for the isolation of exRNA, protocols for detection and quantitation of exRNA, and approaches that can be useful for studies to evaluate potential therapeutic applications. 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, Extracellular RNA: Methods and Protocols aims to ensure successful results in the further study of this vital field. Front Matter ....Pages i-xi Extracellular RNAs: A New Awareness of Old Perspectives (Noah Sadik, Lilian Cruz, Alessandra Gurtner, Rodosthenis S. Rodosthenous, Sophie A. Dusoswa, Olivia Ziegler et al.)....Pages 1-15 Overview of Protocols for Studying Extracellular RNA and Extracellular Vesicles (Julia Small, Sabrina Roy, Roger Alexander, Leonora Balaj)....Pages 17-21 Extracellular RNA Isolation from Cell Culture Supernatant (Sawen Bakr, Bridget Simonson, Kirsty M. Danielson, Saumya Das)....Pages 23-34 Use of a Hollow Fiber Bioreactor to Collect Extracellular Vesicles from Cells in Culture (Irene K. Yan, Neha Shukla, David A. Borrelli, Tushar Patel)....Pages 35-41 Isolation of Extracellular RNA from Serum/Plasma (Justyna Filant, Parham Nejad, Anu Paul, Bridget Simonson, Srimeenakshi Srinivasan, Xuan Zhang et al.)....Pages 43-57 Isolation of Extracellular RNA from Bile (Irene K. Yan, Valentin X. Berdah, Tushar Patel)....Pages 59-67 Cushioned–Density Gradient Ultracentrifugation (C-DGUC): A Refined and High Performance Method for the Isolation, Characterization, and Use of Exosomes (Kang Li, David K. Wong, King Yeung Hong, Robert L. Raffai)....Pages 69-83 Magnetic Particle-Based Immunoprecipitation of Nanoscale Extracellular Vesicles from Biofluids (Pete Heinzelman)....Pages 85-107 Enrichment of Extracellular Vesicle Subpopulations Via Affinity Chromatography (Michelle E. Hung, Stephen B. Lenzini, Devin M. Stranford, Joshua N. Leonard)....Pages 109-124 Detection and Analysis of Non-vesicular Extracellular RNA (Juan Pablo Tosar, Alfonso Cayota)....Pages 125-137 Isolation of Plasma Lipoproteins as a Source of Extracellular RNA (Kang Li, David K. Wong, Fu Sang Luk, Roy Y. Kim, Robert L. Raffai)....Pages 139-153 Droplet Digital PCR for Quantitation of Extracellular RNA (Irene K. Yan, Rishabh Lohray, Tushar Patel)....Pages 155-162 Preparation of Small RNA NGS Libraries from Biofluids (Alton Etheridge, Kai Wang, David Baxter, David Galas)....Pages 163-175 Multiplexed Detection and Quantitation of Extracellular Vesicle RNA Expression Using NanoString (Neha Shukla, Irene K. Yan, Tushar Patel)....Pages 177-185 Milk-derived Extracellular Vesicles for Therapeutic Delivery of Small Interfering RNAs (Akiko Matsuda, Tushar Patel)....Pages 187-197 Loading of Extracellular Vesicles with Hydrophobically Modified siRNAs (Marie-Cecile Didiot, Reka A. Haraszti, Neil Aronin, Anastasia Khvorova)....Pages 199-214 Back Matter ....Pages 215-216