Antiviral Rnai

In this book, expert RNAi specialists from around the world have teamed up to produce a timely and thought-provoking review of the area. The two central themes are: 1) the latest findings on RNAi-virus interactions and 2) progress in the development of RNAi-based antiviral therapeutics. A number of chapters explain general concepts concerned with the role of RNAi in natural antiviral defense mechanisms, other chapters discuss how to improve the efficacy and safety of RNAi-based antiviral drugs, while others describe how this technology is being developed as a new therapeutic tool for fighting.

Viruses and RNAi share an intricate relationship at many levels. RNAi is an important antiviral defense mechanism in plants and invertebrates, microRNAs – of viral or cellular origin – affect many aspects of virus biology, and replication of many, if not all, mammalian viruses can be suppressed by RNAi. Antiviral RNAi: Concepts, Methods, and Applications provides a collection of protocols for the analysis of viral small RNAs and natural antiviral RNAi responses as well as for the development and optimization of RNAi-based antiviral drugs. As RNAi is a central regulatory mechanism in the cell, the methods in this volume can also be applied out of the context of a virus infection. Divided into five convenient parts, this detailed volume reviews important basic concepts in the field of antiviral RNAi, provides experimental and bio-informatic tools for the analysis of small silencing RNAs, covers methods to biochemically dissect RNAi-based antiviral defense and viral counter-defense mechanisms, describes methods for the design, expression, and delivery of therapeutic antiviral siRNAs, and finally presents genome-wide RNAi approaches for the identification of factors involved in virus replication. Written in the highly successful Methods in Molecular BiologyTM series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Antiviral RNAi: Concepts, Methods, and Applications serves as an ideal guide for both novice and experienced researchers alike striving to dissect the role of RNAi in the viral life cycle or to further boost the development of novel therapeutics and experimental tools based on RNAi technology.

Studies related to pathogen-mediated virus resistance in plants were instrumental in providing some of the historical observations which ultimately led to the vital discovery of double-stranded RNA (dsRNA)-induced gene silencing or RNA interference (RNAi), which has since revolutionized research on plant-virus interactions. In Antiviral Resistance in Plants: Methods and Protocols, expert researchers in the field detail many of the methods which are now commonly used to study the phenomenon of RNA silencing in relation to viral infections of plants. These include methods and techniques for the isolation and quantitative/qualitative analyses of plant small 21-24 nucleotide RNAs such as small interfering RNAs (siRNAs) and microRNAs (miRNAs) as well as the analysis and manipulation of virus-induced gene silencing (VIGS) in both monocotyledonous and dicotyledenous plants and the use of hairpin RNA (hpRNA) transgenes. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Antiviral Resistance in Plants: Methods and Protocols seeks to aid scientists in the further study of this crucially important botanical trait.

Recent research has focused attention on the importance of intrinsic antiviral immunity, i.e. immunity mediated by factors that are constitutively expressed in many cells. In this volume, leading experts provide a comprehensive overview of this relatively new and rapidly evolving field. They cover intrinsic proteinaceous antiviral immune effectors, such as the APOBEC3 and TRIM protein families as well as Tetherin and SAMHD1, which were initially discovered by researchers studying HIV-1. Furthermore, the role of RNA interference in antiviral defense in plants and invertebrates, as well as the interplay between microRNAs and viruses in mammalian cells, are analysed. One chapter discusses how intrinsic immunity and viral countermeasures to intrinsic immune effectors drive both pathogen and host evolution, and finally the emerging evidence that DNA damage response proteins restrict infection by DNA viruses is highlighted.

This is the first comprehensive book on human/animal gene responses to RNA viral infections, including prevalent, emerging and re-emerging RNA viruses such as HIV, SARS-CoV, West Nile virus, influenza virus and many others. Human gene responses are reviewed by leading virologists worldwide in the following aspects: (i) the altered gene expression profiles at the transcriptional and translational levels detected with cutting-edge technologies such as cDNA microarray and proteomics; (ii) host innate and adapted immune responses to viral replication in target organs; (iii) virus-activated signal transduction pathways in cell survival, apoptosis and autophagosomal pathways; and (iv) the small interfering RNA/microRNA-mediated gene silencing pathway, a recently characterized new host defense mechanism against viral infection.

The discovery of Toll-like receptors (TLRs) in the late 1990s ushered in a new age of discovery for innate immunity. The importance of TLRs for immunology and biomedical research was recognized with the Nobel Prize for Medicine or Physiology in 2011. The prize was shared by three scientists: Ralph Steinman (for the discovery of dendritic cells, whi

A collection of readily reproducible methods for the design, preparation, and use of RNAs for silencing gene expression in cells and organisms. The techniques range widely and include methods addressing the biochemical aspects of the silencing machinery, RNA silencing in non-mammalian organisms, and the in vivo delivery of siRNAs and silencing vectors. There are also techniques for designing, preparing, and using RNAs to silence gene expression, for fine-tuning regulation by targeting specific isoforms of a given gene, and for the study and use of microRNAs. The protocols follow the successful Methods in Molecular BiologyTM series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.