4.8 Article

MiMIC: a highly versatile transposon insertion resource for engineering Drosophila melanogaster genes

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NATURE METHODS
卷 8, 期 9, 页码 737-U80

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NATURE PUBLISHING GROUP
DOI: 10.1038/nmeth.1662

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  1. US National Institutes of Health [2R01 GM067858, T32 GM07526-33]
  2. Howard Hughes Medical Institute

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We demonstrate the versatility of a collection of insertions of the transposon Minos-mediated integration cassette (MiMIC), in Drosophila melanogaster. MiMIC contains a gene-trap cassette and the yellow(+) marker flanked by two inverted bacteriophage Phi C31 integrase attP sites. MiMIC integrates almost at random in the genome to create sites for DNA manipulation. The attP sites allow the replacement of the intervening sequence of the transposon with any other sequence through recombinase-mediated cassette exchange (RMCE). We can revert insertions that function as gene traps and cause mutant phenotypes to revert to wild type by RMCE and modify insertions to control GAL4 or QF overexpression systems or perform lineage analysis using the Flp recombinase system. Insertions in coding introns can be exchanged with protein-tag cassettes to create fusion proteins to follow protein expression and perform biochemical experiments. The applications of MiMIC vastly extend the D. melanogaster toolkit.

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