Expanding the genetic toolkit of Tribolium castaneum
| Autor: | Dylan J. Siniard, Gabriel E. Zentner, Kaitlin K. Doucette, Johnathan C. Rylee, Andrew C. Zelhof |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Life Cycles Gene Expression lcsh:Medicine Synthetic Genome Editing Genetic analysis Genome Engineering 0302 clinical medicine Larvae Beetles Genes Reporter Gene expression Invertebrate Genomics Gene Order CRISPR Transgenes Promoter Regions Genetic lcsh:Science 0303 health sciences Tribolium Multidisciplinary Fluorescent reporter biology Drosophila Melanogaster Crispr Eukaryota Gene targeting Genomics Animal Models Insects Phenotype Experimental Organism Systems Genetic Techniques Gene Targeting Engineering and Technology Synthetic Biology Drosophila Research Article Biotechnology animal structures Arthropoda Transgene Genetic Vectors Bioengineering Computational biology Research and Analysis Methods 03 medical and health sciences Model Organisms Ribosomal protein Genetics Animals Gene Disruption Red flour beetle Gene 030304 developmental biology fungi lcsh:R Organisms Biology and Life Sciences Synthetic Genomics biology.organism_classification Invertebrates 030104 developmental biology Synthetic Bioengineering Animal Genomics Genetic Loci Genetic marker lcsh:Q Cellular Morphology CRISPR-Cas Systems 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | PLoS ONE, Vol 13, Iss 4, p e0195977 (2018) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | The red flour beetle, Tribolium castaneum, is an important model insect and agricultural pest.However, many standard genetic tools are lacking or underdeveloped in this system. Here, we present a set of new reagents to augment existing Tribolium genetic tools. We demonstrate a new GAL4 driver line that employs the promoter of a ribosomal protein gene to drive expression of a UAS responder in the fat body. We also present a novel dual fluorescent reporter that labels cell membranes and nuclei with different fluorophores for the analysis of cellular morphology. This approach also demonstrates the functionality of the viral T2A peptide for bicistronic gene expression in Tribolium. To facilitate classical genetic analysis, we created lines with visible genetic markers by CRISPR-mediated disruption of the yellow and ebony body color loci with a cassette carrying an attP site, enabling future φC31-mediated integration. Together,the reagents presented here will facilitate more robust genetic analysis in Tribolium and serve as a blueprint for the further development of this powerful model’s genetic toolkit. |
| Databáze: | OpenAIRE |
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