5
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      A transgenic female killing system for the genetic control of Drosophila suzukii

      research-article

      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

          The spotted wing Drosophila ( Drosophila suzukii) is an invasive pest of soft-skinned fruit crops. It is rapidly transmitted in Europe and North America, causing widespread agricultural losses. Genetic control strategies such as the sterile insect technique (SIT) have been proposed as environment-friendly and species-restricted approaches for this pest. However, females are inefficient agents in SIT programs. Here we report a conditional female-killing (FK) strategy based on the tetracycline-off system. We assembled sixteen genetic constructs for testing in vitro and in vivo. Twenty-four independent transgenic strains of D. suzukii were generated and tested for female-specific lethality. The strongest FK effect in the absence of tetracycline was achieved by the construct containing D. suzukii nullo promoter for early gene expression, D. suzukii pro-apoptotic gene hid Ala4 for lethality, and the transformer gene intron from the Mediterranean fruit fly Ceratitis capitata for female-specific splicing. One strain carrying this construct eliminated 100% of the female offspring during embryogenesis and produced only males. However, homozygous females from these FK strains were not viable on a tetracycline-supplemented diet, possibly due to the basal expression of hid Ala4 . Potential improvements to the gene constructs and the use of such FK strains in an SIT program are discussed.

          Related collections

          Most cited references52

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

          Improved splice site detection in Genie.

          We present an improved splice site predictor for the genefinding program Genie. Genie is based on a generalized Hidden Markov Model (GHMM) that describes the grammar of a legal parse of a multi-exon gene in a DNA sequence. In Genie, probabilities are estimated for gene features by using dynamic programming to combine information from multiple content and signal sensors, including sensors that integrate matches to homologous sequences from a database. One of the hardest problems in genefinding is to determine the complete gene structure correctly. The splice site sensors are the key signal sensors that address this problem. We replaced the existing splice site sensors in Genie with two novel neural networks based on dinucleotide frequencies. Using these novel sensors, Genie shows significant improvements in the sensitivity and specificity of gene structure identification. Experimental results in tests using a standard set of annotated genes showed that Genie identified 86% of coding nucleotides correctly with a specificity of 85%, versus 80% and 84% in the older system. In further splice site experiments, we also looked at correlations between splice site scores and intron and exon lengths, as well as at the effect of distance to the nearest splice site on false positive rates.
            Bookmark
            • Record: found
            • Abstract: not found
            • Article: not found

            Invasion biology of spotted wing Drosophila (Drosophila suzukii): a global perspective and future priorities

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

              Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity.

              Regulatory elements that control tetracycline resistance in Escherichia coli were previously converted into highly specific transcription regulation systems that function in a wide variety of eukaryotic cells. One tetracycline repressor (TetR) mutant gave rise to rtTA, a tetracycline-controlled transactivator that requires doxycycline (Dox) for binding to tet operators and thus for the activation of P(tet) promoters. Despite the intriguing properties of rtTA, its use was limited, particularly in transgenic animals, because of its relatively inefficient inducibility by doxycycline in some organs, its instability, and its residual affinity to tetO in absence of Dox, leading to elevated background activities of the target promoter. To remove these limitations, we have mutagenized tTA DNA and selected in Saccharomyces cerevisiae for rtTA mutants with reduced basal activity and increased Dox sensitivity. Five new rtTAs were identified, of which two have greatly improved properties. The most promising new transactivator, rtTA2(S)-M2, functions at a 10-fold lower Dox concentration than rtTA, is more stable in eukaryotic cells, and causes no background expression in the absence of Dox. The coding sequences of the new reverse TetR mutants fused to minimal activation domains were optimized for expression in human cells and synthesized. The resulting transactivators allow stringent regulation of target genes over a range of 4 to 5 orders of magnitude in stably transfected HeLa cells. These rtTA versions combine tightness of expression control with a broad regulatory range, as previously shown for the widely applied tTA.
                Bookmark

                Author and article information

                Contributors
                Ying.Yan@agrar.uni-giessen.de
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                21 June 2021
                21 June 2021
                2021
                : 11
                : 12938
                Affiliations
                [1 ]GRID grid.8664.c, ISNI 0000 0001 2165 8627, Institute for Insect Biotechnology, Department of Insect Biotechnology in Plant Protection, , Justus-Liebig-University Giessen, ; Winchesterstraße 2, 35394 Giessen, Germany
                [2 ]GRID grid.418010.c, ISNI 0000 0004 0573 9904, Fraunhofer Institute for Molecular Biology and Applied Ecology IME, ; Winchesterstraße 2, 35394 Giessen, Germany
                Article
                91938
                10.1038/s41598-021-91938-1
                8217240
                34155227
                e652bd54-1d51-4728-bc20-b6414320adee
                © The Author(s) 2021

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 9 March 2021
                : 2 June 2021
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100003185, Fraunhofer-Gesellschaft;
                Award ID: Applications for population control of D. suzukii
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100004493, International Atomic Energy Agency;
                Funded by: Justus-Liebig-Universität Gießen (3114)
                Categories
                Article
                Custom metadata
                © The Author(s) 2021

                Uncategorized
                synthetic biology,genetic engineering,molecular engineering,entomology
                Uncategorized
                synthetic biology, genetic engineering, molecular engineering, entomology

                Comments

                Comment on this article