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Double-Stranded RNA

Methods and Protocols

  • Book
  • © 2024

Overview

Editors:
  1. Xiaofei Cheng

    1. College of Plant Protection/Key Laboratory of Germplasm Enhancement, Physiology and Ecology of Food Crops in Cold Region of Chinese Education Ministry, Northeast Agricultural University, Harbin, China

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    You can also search for this editor in PubMed   Google Scholar

  2. Guanwei Wu

    1. Institute of Plant Virology, Ningbo University, Ningbo City, China

    View editor publications

    You can also search for this editor in PubMed   Google Scholar

  • Includes cutting-edge techniques
  • Provides step-by-step detail essential for reproducible results
  • Contains key implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2771)

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About this book

This detailed volume examines classical and cutting-edge methods involving double-stranded RNA (dsRNA), specifically regarding isolation, visualization, characterization, production, and application. Many protocols, such as co-immunoprecipitation-based isolation of double-stranded RNA-associated protein complexes, identification of mycoviruses by dsRNA extraction, application of dsRNA for fungi disease management (Sclerotinia sclerotiorum and Botrytis cin), and production of double-stranded RNA in plants by plant viral vectors for gene silencing, can also be easily adapted for identification of viruses from other organisms, control of other pathogens, and fundamental research. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips for troubleshooting and avoiding known pitfalls. 

Reliable and practical, Double-Stranded RNA: Methods and Protocols serves as an ideal reference book for students and researchers who work with dsRNA.

Keywords

Table of contents (18 protocols)

  1. Front Matter

    Pages i-xi
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  2. Isolation of Double-Stranded RNAs by Lithium Chloride Fractionation

    • Mengzhu Chai, Xinyue Ma, Shuaifa Chang, Xiaofei Cheng
    Pages 1-5

  3. Detection of dsRNA by Acridine Orange Staining

    • Tingshuai Ma, Yu Zhao, Xiaofei Cheng
    Pages 7-12

  4. Isolation of dsRNA from Plants by Cellulose Chromatography

    • Yameng Luan, Wenqian Fan, Xayvangye Korxeelor, Xiaoyun Wu
    Pages 13-17

  5. Rapid Purification of dsRNA Using Micro-Spin Cellulose Column

    • Yuting Wang, Ziyi Wang, Xiaoyun Wu
    Pages 19-25

  6. Analysis of Virus-Induced Double-Stranded RNA in Living Plant Cells by the dRBFC Assay

    • Ying Zhang, Xinyue Fan, Xiaofei Cheng
    Pages 27-33

  7. Detection of dsRNA with Fluorescence In Situ Hybridization (FISH)

    • Wenling Zhou, Juan Huang, Xianguang Yang, Xiaoming Zhang
    Pages 35-38

  8. Subcellular Colocalization Assay of Host Factors with Viral Replication Complex in the dsRNA Reporter Nicotiana benthamiana

    • Xinxin Fang, Jianping Chen, Fei Yan, Guanwei Wu
    Pages 39-45

  9. Production of Double-Stranded RNA Using the Prokaryotic Promoter-Mediated Bidirectional Transcription

    • Xue Jiang, Kekely Bruno Attiogbe, Yating Guo, Xiaoyun Wu
    Pages 47-55

  10. Inducible Expression of dsRNA in Escherichia coli

    • Saiya Duan, Guangjun Wang
    Pages 57-64

  11. In Vivo Production of dsRNA Using Bacteriophage ϕ6 in Pseudomonas syringae Cit7 Cells

    • Jinguang Yang
    Pages 65-72

  12. Bacteria-Based Double-Stranded RNA Production to Develop Cost-Effective RNA Interference Application for Insect Pest Management

    • Ruobing Guan, Xuexia Miao, Haichao Li
    Pages 73-81

  13. Transiently Induce RNA Silencing in Plants Using a Tobacco Necrosis Virus A (TNV-A)-Based dsRNA Production System

    • Yuanming Zhang, Mengzhu Chai, Xiaofei Cheng, Kai Xu
    Pages 83-89

  14. Co-immunoprecipitation-Based Isolation of Double-Stranded RNA-Associated Protein Complexes in Nicotiana benthamiana

    • Zhaoxing Jia, Jianping Chen, Fei Yan, Guanwei Wu
    Pages 91-98

  15. Analysis of Plant Virus-Induced Immunity by Using Viral-Derived Double-Stranded RNA in Arabidopsis thaliana

    • Penghuan Rui, Jianping Chen, Fei Yan, Guanwei Wu
    Pages 99-110

  16. Identification of Mycoviruses by dsRNA Extraction

    • Yanfei Wang, Clement Nzabanita, Lihua Guo
    Pages 111-118

  17. Production of Double-Stranded RNA in Planta by a Potato Mop-Top Virus (PMTV)-Based Vector for Inducing Gene Silencing

    • Ye Liu, Yanlin Gao, Xiaofei Cheng, Yanju Bai
    Pages 119-126

  18. Application of dsRNA for Fungi Disease Management Sclerotinia sclerotiorum and Botrytis cinerea

    • Tao Tang, Yumeng Wang, Dongdong Niu
    Pages 127-132

  19. Application of dsRNA in the Pine Wood Nematode, Bursaphelenchus xylophilus

    • Chunyu Wang, Kai Guo
    Pages 133-139

  20. Back Matter

    Pages 141-142
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Editors and Affiliations

  • College of Plant Protection/Key Laboratory of Germplasm Enhancement, Physiology and Ecology of Food Crops in Cold Region of Chinese Education Ministry, Northeast Agricultural University, Harbin, China

    Xiaofei Cheng

  • Institute of Plant Virology, Ningbo University, Ningbo City, China

    Guanwei Wu

Bibliographic Information

  • Book Title: Double-Stranded RNA

  • Book Subtitle: Methods and Protocols

  • Editors: Xiaofei Cheng, Guanwei Wu

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-3702-9

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024

  • Hardcover ISBN: 978-1-0716-3701-2Published: 30 January 2024

  • Softcover ISBN: 978-1-0716-3704-3Due: 12 February 2025

  • eBook ISBN: 978-1-0716-3702-9Published: 29 January 2024

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XI, 142

  • Number of Illustrations: 11 b/w illustrations, 20 illustrations in colour

  • Topics: Biochemistry, general, Genetics and Genomics, Virology

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