An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrases

• DOI: 10.1073/pnas.0611511104
• Publication Date: 2007-02-23T01:41:40Z
• Authors: Johannes Bischof; Robert K. Maeda; Monika Hediger; François Karch; Konrad Basler
• Keywords: Transgenesis; Transposase; Integrases

Germ-line transformation via transposable elements is a powerful tool to study gene function in Drosophila melanogaster. However, some inherent characteristics of transposon-mediated transgenesis limit its use for transgene analysis. Here, we circumvent these limitations by optimizing phiC31-based integration system. We generated collection lines with precisely mapped attP sites that allow the insertion transgenes into many different predetermined intergenic locations throughout fly genome. By using regulatory nanos and vasa genes, established endogenous sources phiC31 integrase, eliminating difficulties coinjecting integrase mRNA raising efficiency. Moreover, discriminate between specific rare nonspecific events, white gene-based reconstitution system was enables visual selection precise targeting. Finally, demonstrate our chromosomal can be modified situ, extending their scope while retaining properties as landing sites. The efficiency, ease-of-use, versatility obtained here represents an important advance opens up possibility systematic, high-throughput screening large cDNA sets elements.