1. Extracellular RNAs – A New Awareness of Old Perspectives
Noah Sadik, Lilian Cruz, Alessandra Gurtner, Rodosthenis S. Rodosthenous, Sophie A. Dusoswa, Olivia Ziegler, Thomas Sebastiaan Van Solinge, Zhiyun Wei, Ane Miren Salvador Garicano, Bence Gyorgy, Marike Broekman, and Leonora Balaj
2. Overview of Protocols for Studying Extracellular RNA and Extracellular Vesicles
Julia Small,Roger Alexander, and Leonora Balaj
3. Extracellular RNA Isolation from Cell Culture Supernatant
Sawen Bakr., Bridget Simonson, Kirsty M. Danielson, and Saumya Das
4. Use of a Hollow Fiber Bioreactor to Collect Extracellular Vesicles from Cells in Culture
Irene K. Yan, Neha Shukla, David A. Borrelli, and Tushar Patel
5. Isolation of Extracellular RNA from Serum/Plasma
Justyna Filant, Parham Nejad, Anu Paul, Bridget Simonson, Srimeenakshi Srinivasan, Xuan Zhang, Leonora Balaj, Saumya Das, Roopali Gandhi, Louise C. Laurent,and Anil K. Sood
6. Isolation of Extracellular RNA from Bile
Irene K. Yan, Valentin X. Berdah, and Tushar Patel
7. Cushioned–Density Gradient Ultracentrifugation (C–DGUC): a Refined and High Performance Method for the Isolation, Characterization & Use of Exosomes
Kang Li, David K. Wong, King Yeung Hong, and Robert L. Raffai
8. Magnetic Particle-Based Immunoprecipitation of Nanoscale Extracellular Vesicles from Biofluids
Pete Heinzelman
9. Enrichment of Extracellular Vesicle Subpopulations via Affinity Chromatography
Michelle E. Hung, Stephen B. Lenzini, Devin M. Stranford, and Joshua N. Leonard
10. Detection and Analysis of Non-vesicular Extracellular RNA
Juan Pablo Tosar and Alfonso Cayota
11. Isolation of Plasma Lipoproteins as a Source of Extracellular RNA
Kang Li, David K. Wong, Fu Sang Luk, Roy Y. Kim, and Robert L. Raffai
12. Droplet Digital PCR for Quantitation of Extracellular RNA
Irene K Yan, Rishabh Lohray, and Tushar Patel
13. Preparation of Small RNA NGS Libraries from Biofluids
Alton Etheridge, Kai Wang, David Baxter, and David Galas
14. Multiplexed Detection and Quantitation of Extracellular Vesicle RNA Expression using NanoString
Neha Shukla, Irene K. Yan, and Tushar Patel
15. Milk-derived Extracellular Vesicles for Therapeutic Delivery of Small Interfering RNAs
Akiko Matsuda and Tushar Patel
16. Loading of Extracellular Vesicles with Hydrophobically Modified siRNAs
Marie-Cécile Didiot, Reka A. Haraszti, Neil Aronin, and Anastasia Khvorova
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.