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Genetics. 1999 Jul;152(3):1037-44.

I-SceI endonuclease, a new tool for studying DNA double-strand break repair mechanisms in Drosophila.

Author information

1
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.

Abstract

As a step toward the development of a homologous recombination system in Drosophila, we have developed a methodology to target double-strand breaks (DSBs) to a specific position in the Drosophila genome. This method uses the mitochondrial endonuclease I-SceI that recognizes and cuts an 18-bp restriction site. We find that >6% of the progeny derived from males that carry a marker gene bordered by two I-SceI sites and that express I-SceI in their germ line lose the marker gene. Southern blot analysis and sequencing of the regions surrounding the I-SceI sites revealed that in the majority of the cases, the introduction of DSBs at the I-SceI sites resulted in the complete deletion of the marker gene; the other events were associated with partial deletion of the marker gene. We discuss a number of applications for this novel technique, in particular its use to study DSB repair mechanisms.

PMID:
10388822
PMCID:
PMC1460675
[Indexed for MEDLINE]
Free PMC Article

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