This volume looks at the latest techniques used in the study of Rift Valley Fever Virus (RVFV). The chapters in this book cover topics such as molecular biology, biochemistry, and cell biology approaches that enable the mechanistic study of virus-host interactions; vector competence, detection, and diagnosis; virus inactivation for biosafety purposes; virus and protein production for structural studies; high-throughput screening based on CRISPR-Cas9 technology; single-molecule RNA imaging; and physiological infection models, animal models, and immunology. Written in the high successful Methods in Molecular Biology 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.

Cutting-edge and comprehensive, Rift Valley Fever Virus: Methods and Protocols is a valuable resource for both novice and experts researchers who want to learn more about the important and developing field of RVFV.

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

Inhalt

An Introduction to Rift Valley Fever Virus.- Aedes aegypti Vector Competence Assay for Rift Valley Fever Virus using Artificial Blood Meal.- Isolation of Rift Valley Fever Virus from Field Specimens using Cell Culture Approaches.- Detection and Diagnosis of Rift Valley Fever Virus.- Analysis of Negative-Strand RNA Viruses by RT-qPCR: Rift Valley Fever Virus and Toscana Virus.- Validated Methods for Effective Decontamination and Inactivation of Rift Valley Fever Virus.- Guidelines for In Vitro Production and Quantification of Rift Valley Fever Virus.- Cell-Free Synthesis of Bunyavirales Proteins in View of Their Structural Characterization by Nuclear Magnetic Resonance.- Expression and Purification of RVFV Glycoprotein C for Structural Studies using Baculovirus Expression System.- Next-Generation Sequencing and Proteomics-Enabled Approach for Rapid and High-Throughput Isolation of Virus-Neutralizing Nanobodies.- Use of Single-Domain Antibodies Against the NSm Protein for the Detection of Cells Infected by Rift Valley Fever Virus.- Making Rift Valley Fever Viral Particles Fluorescent.- Identification of RVFV Host Factors using Quantitative Interaction Proteomics.- Genome-Wide CRISPR-Cas9 Screening for the Identification of Host Dependency Factors of Emerging Viruses.- Preparation of Bunyavirus-Infected Cells for Electron Cryo-Tomography.- Imaging Bunyavirus Infections by Transmission Electron Microscopy: Conventional Sample Preparation vs High-Pressure Freezing and Freeze-Substitution.- The Phlebovirus Ribonucleoprotein: An Overview.- Nucleotide Resolution Mapping of Rift Valley Fever Virus Nucleoprotein - Genome RNA Interactions.- Characterization of RVFV Nucleocapsid Protein Binding Sites on RNA by iCLIP-seq.- Yeast Expression and Activity Assay of RVFV Polymerase.- Single-Molecule Visualization of Bunyavirus Genome Segments using Fluorescence In Situ Hybridization.- Single-Cell RNA-Sequencing of RVFV Infection.- Using RNAseq to Uncover Transcriptional and Splicing Differences in Host Cells during Rift Valley Fever Virus Infection.- Quantification of RVFV Specific T Cell Responses in Mice Pre-Immunized with Potential Vaccine Candidates.- Use of Human Macrophages to Study Bunyavirus NSs Functions.- 3D Spheroid and Organoid Models to Study Neuroinfection of RNA Viruses.- Laboratory Animal Models for Rift Valley Fever Virus Disease.- Monitoring RVFV Infection using Bioluminescent Reporter Viruses In Vivo.