title: Highly efficient integration and expression of piggyBac-derived cassettes in the honeybee (Apis mellifera).
abstract: Honeybees (Apis mellifera), which are important pollinators of plants, display remarkable individual behaviors that collectively contribute to the organization of a complex society. Advances in dissecting the complex processes of honeybee behavior have been limited in the recent past due to a lack of genetic manipulation tools. These tools are difficult to apply in honeybees because the unit of reproduction is the colony, and many interesting phenotypes are developmentally specified at later stages. Here, we report highly efficient integration and expression of piggyBac-derived cassettes in the honeybee. We demonstrate that 27 and 20% of queens stably transmitted two different expression cassettes to their offspring, which is a 6- to 30-fold increase in efficiency compared with those generally reported in other insect species. This high efficiency implies that an average beekeeping facility with a limited number of colonies can apply this tool. We demonstrated that the cassette stably and efficiently expressed marker genes in progeny under either an artificial or an endogenous promoter. This evidence of efficient expression encourages the use of this system to inhibit gene functions through RNAi in specific tissues and developmental stages by using various promoters. We also showed that the transgenic marker could be used to select transgenic offspring to be employed to facilitate the building of transgenic colonies via the haploid males. We present here the first to our knowledge genetic engineering tool that will efficiently allow for the systematic detection and better understanding of processes underlying the biology of honeybees.
authors.affiliation: Department of Evolutionary Genetics, Heinrich Heine University Duesseldorf, D-40225 Duesseldorf, Germany christina-schulte@gmx.de.
authors.forename: Christina
authors.initials: C
authors.lastname: Schulte
authors.affiliation: Department of Evolutionary Genetics, Heinrich Heine University Duesseldorf, D-40225 Duesseldorf, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Theilenberg
authors.affiliation: Department of Evolutionary Genetics, Heinrich Heine University Duesseldorf, D-40225 Duesseldorf, Germany.
authors.forename: Marion
authors.initials: M
authors.lastname: Müller-Borg
authors.affiliation: Department of Evolutionary Genetics, Heinrich Heine University Duesseldorf, D-40225 Duesseldorf, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Gempe
authors.affiliation: Department of Evolutionary Genetics, Heinrich Heine University Duesseldorf, D-40225 Duesseldorf, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Beye
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D001516:Bees / Q000235:genetics*; D001522:Behavior, Animal; D004251:DNA Transposable Elements; D005260:Female; D015723:Gene Library; D017930:Genes, Reporter; D005818:Genetic Engineering; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000737:chemistry; D008297:Male; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D034622:RNA Interference; D019076:Transgenes; D019895:Transposases / Q000378:metabolism; D014231:Tribolium
chemical_list: D004251:DNA Transposable Elements; D049452:Green Fluorescent Proteins; D019895:Transposases
keywords: social insect transgene; transformation; transposon