[PDF] Antiviral Rnai eBook

Author : Ronald P. van Rij
Publisher : Humana Press
Page : 0 pages
File Size : 27,4 MB
Release : 2011-03-08
Category : Medical
ISBN : 9781617790362

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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.

Author : Ronald P. van Rij
Publisher : Humana Press
Page : 425 pages
File Size : 37,49 MB
Release : 2016-08-23
Category : Medical
ISBN : 9781493958252

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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.

Author : Jing Zhong
Publisher :
Page : 129 pages
File Size : 19,51 MB
Release : 2014
Category : Caenorhabditis elegans
ISBN : 9781321321029

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RNA interference (RNAi) acts as an antiviral defense mechanism in fungi, plants, nematodes, insects, and mammals. In antiviral RNAi, virus-specific double-stranded RNA is processed into small interfering RNAs (siRNAs) to guide specific viral RNA degradation by the RNAi machinery. Although antiviral RNAi is non cell-autonomous in plants, it is unknown if antiviral RNAi is also systemic in animals. In this dissertation, I characterized the nematode Caenorhabditis elegans mutants defective in systemic RNAi in their antiviral RNAi response induced by either the replication of a Flock house virus-derived replicon or the infection of Orsay virus. The results from these genetic studies provided evidence for the first time to support an antiviral function of systemic RNAi in animals. Comparison of the population of viral siRNAs by deep sequencing further revealed that C. elegans mutants with strong defects in systemic antiviral RNAi were all partially defective in the biogenesis of the viral secondary siRNAs. A possible role for the viral siRNAs in systemic antiviral RNAi is discussed.

Author : Benjamin Obadia
Publisher :
Page : 156 pages
File Size : 16,11 MB
Release : 2013
Category :
ISBN :

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Insects have successfully adapted to an incredible variety of environments where they co-exist with diverse microorganisms (e.g., fungi, bacteria, viruses). Consequently, insect defense mechanisms have been shaped over their evolution to generate a versatile immune system, allowing them to better survive infection, but also to better spread infection to others. Although insect-borne diseases have caused severe threats to humans since recorded history, studies had mainly focused on the dissection and understanding of insect-bacterium interactions while insect-virus interactions have remained poorly characterized. A few years ago, the discovery of RNA interference (RNAi) as an antiviral immune mechanism opened new perspectives on understanding insect immunity, which may potentially lead to the control of insect-borne viruses. RNAi is naturally triggered by virus-derived double-stranded (ds) RNA molecules and gives the insect a sequence-specific way of controlling viral replication. In insects, the antiviral RNAi response may also be mounted after artificial immunization with dsRNA, and evidence shows that a systemic protective state is established against viruses. Using the insect model Drosophila melanogaster, the present work inquires on the capacity of dsRNA to generate such a systemic silencing response against RNA viruses and focuses on the fate of dsRNA once in the insect organism. We provide evidence that the intestinal epithelium is the principal tissue involved in dsRNA uptake from both environmental and systemic media. The antiviral immune property of the midgut mediated by RNA interference is also debated.

Author : John M. Watson
Publisher : Humana Press
Page : 380 pages
File Size : 48,19 MB
Release : 2012-06-12
Category : Science
ISBN : 9781617798832

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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.

Author : Bryan R. Cullen
Publisher : Springer Science & Business Media
Page : 265 pages
File Size : 12,10 MB
Release : 2013-05-17
Category : Medical
ISBN : 3642377653

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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.

Author : Patrick Arbuthnot
Publisher : Academic Press
Page : 391 pages
File Size : 41,54 MB
Release : 2015-06-01
Category : Science
ISBN : 0124114520

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Gene Therapy for Viral Infections provides a comprehensive review of the broader field of nucleic acid and its use in treating viral infections. The text bridges the gap between basic science and important clinical applications of the technology, providing a systematic, integrated review of the advances in nucleic acid-based antiviral drugs and the potential advantages of new technologies over current treatment options. Coverage begins with the fundamentals, exploring varying topics, including harnessing RNAi to silence viral gene expression, antiviral gene editing, viral gene therapy vectors, and non-viral vectors. Subsequent sections include detailed coverage of the developing use of gene therapy for the treatment of specific infections, the principles of rational design of antivirals, and the hurdles that currently face the further advancement of gene therapy technology. Provides coverage of gene therapy for a variety of infections, including HBV, HCV, HIV, hemorrhagic fever viruses, and respiratory and other viral infections Bridges the gap between the basic science and the important medical applications of this technology Features a broad approach to the topic, including an essential overview and the applications of gene therapy, synthetic RNA, and other antiviral strategies that involve nucleic acid engineering Presents perspectives on the future use of nucleic acids as a novel class of antiviral drugs Arms the reader with the cutting-edge information needed to stay abreast of this developing field

Author : Jinfeng Lu
Publisher :
Page : 133 pages
File Size : 48,96 MB
Release : 2015
Category : Antiviral agents
ISBN : 9781339467412

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Diverse eukaryotic hosts produce virus-derived small interfering RNAs (siRNAs) to direct antiviral immunity by RNA interference (RNAi). However, it has been controversial in the past decade whether the mammalian RNAi pathway has a natural antiviral function. In this dissertation, I demonstrate the production of canonical virus-derived siRNAs processed by endoribonuclease Dicer from viral double-stranded RNA (dsRNA) precursors in cultured hamster and human somatic cells and in mice infected by two distinct RNA viruses after their cognate viral suppressor of RNA silencing (VSR) is rendered inactive. These mammalian viral siRNAs are predominantly 22 nucleotides long and contain mostly uridine as the 5'-terminal nucleotide, and are therefore similar to mammalian microRNAs. I show that canonical viral siRNAs are loaded into mouse and human Argonaute proteins at high levels in the infected cells. Further analysis reveals that the VSR protein NS1 of Influenza A virus inhibits the biogenesis of viral siRNAs whereas the VSR protein B2 of Nodamura virus suppresses both the biogenesis and the Argonaute loading of viral siRNAs during infection. The results together demonstrate that the antiviral RNAi response is conserved in mammals as found in fungi, plants, insects, and nematodes.

Author : Gordon Carmichael
Publisher : Springer Science & Business Media
Page : 343 pages
File Size : 29,91 MB
Release : 2008-02-04
Category : Science
ISBN : 1592599354

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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.