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Mobile Genetic Elements

Volume 859 of the series Methods in Molecular Biology pp 203-228

Date:

Genome-Wide Manipulations of Drosophila melanogaster with Transposons, Flp Recombinase, and ΦC31 Integrase

  • Koen J. T. VenkenAffiliated withDepartment of Molecular and Human Genetics, Howard Hughes Medical Institute, Baylor College of Medicine Email author 
  • , Hugo J. BellenAffiliated withDepartment of Molecular and Human Genetics, Howard Hughes Medical Institute Program in Developmental Biology, Baylor College of Medicine Email author 

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Abstract

Transposable elements, the Flp recombinase, and the ΦC31 integrase are used in Drosophila melanogaster for numerous genome-wide manipulations. Often, their use is combined in a synergistic fashion to alter and engineer the fruit fly genome. Transposons are the foundation for all transgenic technologies in flies and hence almost all innovations in the fruit fly. They have been instrumental in the generation of genome-wide collections of insertions for gene disruption and manipulation. Many important transgenic strains of these collections are available from public repositories. The Flp protein is the most widely used recombinase to induce mitotic clones to study individual gene function. However, Flp has also been used to generate chromosome- and genome-wide collections of precise deletions, inversions, and duplications. Similarly, transposons that contain attP attachment sites for the ΦC31 integrase can be used for numerous applications. This integrase was incorporated into a transgenesis system that allows the integration of small to very large DNA fragments that can be easily manipulated through recombineering. This system allowed the creation of genomic DNA libraries for genome-wide gene manipulations and X chromosome duplications. Moreover, the attP sites are being used to create libraries of tens of thousands of RNAi constructs and tissue-specific GAL4 lines. This chapter focuses on genome-wide applications of transposons, Flp recombinase, and ΦC31 integrase that greatly facilitate experimental manipulation of Drosophila.

Key words

P element transposon piggyBac transposon Minos transposon Flp recombinase ΦC31 integrase Deletion Duplication Recombineering GAL4/UAS RNAi