Abstract
Transposons such as P elements are routinely used to stably transfer exogenous DNA (transgenes) into the Drosophila genome. Transgene insertion events, however, are essentially random and are subject to 'position effects' from nearby endogenous regulatory elements. Here we describe a microinjection-based system that uses Cre-mediated recombination to insert transgenes into precise genomic 'landing sites'. The system is simple and efficient, and will permit precise comparisons between multiple transgenic constructs.
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Acknowledgements
We thank M. Siegal for the pMLS104 Cre helper plasmid used in these experiments, V. Vasisht for plasmids containing loxP sequences, D. Papatsenko for loxP-y-loxP transgenic flies, M. Wernet for advice on the inverted PCR protocol, and M. Siegal and P. Struffi for comments on the manuscript. L.K. was supported by the 2004 Howard Hughes Medical Institute summer program for high school teachers. This work was supported by grant RO1 GM 51946 from the National Institutes of Health, and conducted in a facility constructed with support from Research Facilities Improvement Grant C06 RR-15518-01 from the National Center for Research Resources, National Institutes of Health.
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Supplementary information
Supplementary Table 1
Cre-mediated RMCE in transgenic Drosophila. (PDF 94 kb)
Supplementary Table 2
Transgene excision tests. (PDF 96 kb)
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Oberstein, A., Pare, A., Kaplan, L. et al. Site-specific transgenesis by Cre-mediated recombination in Drosophila. Nat Methods 2, 583–585 (2005). https://doi.org/10.1038/nmeth775
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DOI: https://doi.org/10.1038/nmeth775
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