Blog
About

20
views
0
recommends
+1 Recommend
1 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      Reduced stability and intracellular transport of dsRNA contribute to poor RNAi response in lepidopteran insects

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          ABSTRACT

          RNA interference (RNAi) has become a widely used reverse genetic tool to study gene function in eukaryotic organisms and is being developed as a technology for insect pest management. The efficiency of RNAi varies among organisms. Insects from different orders also display differential efficiency of RNAi, ranging from highly efficient (coleopterans) to very low efficient (lepidopterans). We investigated the reasons for varying RNAi efficiency between lepidopteran and coleopteran cell lines and also between the Colorado potato beetle , Leptinotarsa decemlineata and tobacco budworm , Heliothis virescens. The dsRNA either injected or fed was degraded faster in H. virescens than in L. decemlineata. Both lepidopteran and coleopteran cell lines and tissues efficiently took up the dsRNA. Interestingly, the dsRNA administered to coleopteran cell lines and tissues was taken up and processed to siRNA whereas the dsRNA was taken up by lepidopteran cell lines and tissues but no siRNA was detected in the total RNA isolated from these cell lines and tissues. The data included in this paper showed that the degradation and intracellular transport of dsRNA are the major factors responsible for reduced RNAi efficiency in lepidopteran insects.

          Related collections

          Most cited references 56

          • Record: found
          • Abstract: found
          • Article: not found

          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            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.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              RNA interference.

              A conserved biological response to double-stranded RNA, known variously as RNA interference (RNAi) or post-transcriptional gene silencing, mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes. RNAi has been cultivated as a means to manipulate gene expression experimentally and to probe gene function on a whole-genome scale.
                Bookmark

                Author and article information

                Journal
                RNA Biol
                RNA Biol
                KRNB
                krnb20
                RNA Biology
                Taylor & Francis
                1547-6286
                1555-8584
                July 2016
                31 May 2016
                31 May 2016
                : 13
                : 7
                : 656-669
                Affiliations
                [a ]Department of Entomology, College of Agriculture, Food and Environment, Agriculture Science Center North, University of Kentucky , Lexington, KY, USA
                [b ]Agricultural Biotechnology Research and Development, DuPont Pioneer , Johnston, IA, USA
                [c ]DowAgrosciences LLC , Indianapolis, Indiana, USA
                Author notes
                Subba Reddy Palli rpalli@ 123456email.uky.edu , rpalli@ 123456email.uky.edu Department of Entomology, Ag. Science N. University of Kentucky Lexington , KY, USA

                Supplemental data for this article can be accessed on the publisher's website.

                Article
                1191728
                10.1080/15476286.2016.1191728
                4962799
                27245473
                © 2016 The Author(s). Published with license by Taylor & Francis Group, LLC

                This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

                Page count
                Figures: 9, Tables: 0, References: 58, Pages: 14
                Product
                Categories
                Research Paper

                Molecular biology

                double-stranded rna, coleoptera, lepidoptera, rna interference

                Comments

                Comment on this article