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RNAi Efficiency, Systemic Properties, and Novel Delivery Methods for Pest Insect Control: What We Know So Far

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      In recent years, the research on the potential of using RNA interference (RNAi) to suppress crop pests has made an outstanding growth. However, given the variability of RNAi efficiency that is observed in many insects, the development of novel approaches toward insect pest management using RNAi requires first to unravel factors behind the efficiency of dsRNA-mediated gene silencing. In this review, we explore essential implications and possibilities to increase RNAi efficiency by delivery of dsRNA through non-transformative methods. We discuss factors influencing the RNAi mechanism in insects and systemic properties of dsRNA. Finally, novel strategies to deliver dsRNA are discussed, including delivery by symbionts, plant viruses, trunk injections, root soaking, and transplastomic plants.

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      Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.

       S Kostas,  A Fire,  S Xu (1998)
      Experimental introduction of RNA into cells can be used in certain biological systems to interfere with the function of an endogenous gene. Such effects have been proposed to result from a simple antisense mechanism that depends on hybridization between the injected RNA and endogenous messenger RNA transcripts. RNA interference has been used in the nematode Caenorhabditis elegans to manipulate gene expression. Here we investigate the requirements for structure and delivery of the interfering RNA. To our surprise, we found that double-stranded RNA was substantially more effective at producing interference than was either strand individually. After injection into adult animals, purified single strands had at most a modest effect, whereas double-stranded mixtures caused potent and specific interference. The effects of this interference were evident in both the injected animals and their progeny. Only a few molecules of injected double-stranded RNA were required per affected cell, arguing against stochiometric interference with endogenous mRNA and suggesting that there could be a catalytic or amplification component in the interference process.
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        Systematic functional analysis of the Caenorhabditis elegans genome using RNAi.

        A principal challenge currently facing biologists is how to connect the complete DNA sequence of an organism to its development and behaviour. Large-scale targeted-deletions have been successful in defining gene functions in the single-celled yeast Saccharomyces cerevisiae, but comparable analyses have yet to be performed in an animal. Here we describe the use of RNA interference to inhibit the function of approximately 86% of the 19,427 predicted genes of C. elegans. We identified mutant phenotypes for 1,722 genes, about two-thirds of which were not previously associated with a phenotype. We find that genes of similar functions are clustered in distinct, multi-megabase regions of individual chromosomes; genes in these regions tend to share transcriptional profiles. Our resulting data set and reusable RNAi library of 16,757 bacterial clones will facilitate systematic analyses of the connections among gene sequence, chromosomal location and gene function in C. elegans.
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          RNA interference is mediated by 21- and 22-nucleotide RNAs.

          Double-stranded RNA (dsRNA) induces sequence-specific posttranscriptional gene silencing in many organisms by a process known as RNA interference (RNAi). Using a Drosophila in vitro system, we demonstrate that 21- and 22-nt RNA fragments are the sequence-specific mediators of RNAi. The short interfering RNAs (siRNAs) are generated by an RNase III-like processing reaction from long dsRNA. Chemically synthesized siRNA duplexes with overhanging 3' ends mediate efficient target RNA cleavage in the lysate, and the cleavage site is located near the center of the region spanned by the guiding siRNA. Furthermore, we provide evidence that the direction of dsRNA processing determines whether sense or antisense target RNA can be cleaved by the siRNA-protein complex.

            Author and article information

            1Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University Gent, Belgium
            2Department of Crop Protection, Molecular Entomology, Federal University of Pelotas Pelotas, Brazil
            Author notes

            Edited by: Arash Zibaee, University of Gilan, Iran

            Reviewed by: Andrew Dacks, West Virginia University, USA; Takashi Koyama, Instituto Gulbenkian de Ciência, Portugal

            *Correspondence: Moises J. Zotti moises.zotti@

            This article was submitted to Invertebrate Physiology, a section of the journal Frontiers in Physiology

            Front Physiol
            Front Physiol
            Front. Physiol.
            Frontiers in Physiology
            Frontiers Media S.A.
            17 November 2016
            : 7
            27909411 5112363 10.3389/fphys.2016.00553
            Copyright © 2016 Joga, Zotti, Smagghe and Christiaens.

            This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

            Figures: 2, Tables: 1, Equations: 0, References: 135, Pages: 14, Words: 11996
            Funded by: Fonds Wetenschappelijk Onderzoek 10.13039/501100003130
            Funded by: Bijzonder Onderzoeksfonds 10.13039/501100007229
            Funded by: Agentschap voor Innovatie door Wetenschap en Technologie 10.13039/501100003132

            Anatomy & Physiology

            pest control, uptake, delivery, systemic rnai, rna interference (rnai)


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