Cancer Institute A national cancer institute
designated cancer center

Roeland Nusse

Publication Details

  • Creating transgenic Drosophila by microinjecting the site-specific phi C31 integrase mRNA and a transgene-containing donor plasmid NATURE PROTOCOLS Fish, M. P., Groth, A. C., Calos, M. P., Nusse, R. 2007; 2 (10): 2325-2331

    Abstract:

    We describe a microinjection-based phiC31 integrase mRNA-mediated method for creating transgenic Drosophila strains. This approach is more efficient than traditional methods and ensures that the transgene is targeted to a precise genomic position. The method involves targeting integration of an exogenous plasmid (containing the transgene and sequences to facilitate integration) to a preplaced recipient site in the Drosophila genome. The plasmid is coinjected into embryos with mRNA encoding the phiC31 integrase, the enzyme that catalyzes the integration reaction. Using the protocol described here, transgenic lines can be established from, on average, 46% of fertile adults obtained after injection, and all integrations should be targeted to the chosen genomic insertion site. The whole procedure, from injection to established transgenic stocks, can be completed in three generations (approximately 1 month) and can be adapted for other types of transgenesis and mRNA injections in Drosophila.

    View details for DOI 10.1038/nprot.2007.328

    View details for Web of Science ID 000253139900004

    View details for PubMedID 17947973

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