Expression of I-CreI endonuclease generates deletions within the rDNA of Drosophila

Silvana Paredes, Keith A. Maggert

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

The rDNA arrays in Drosophila contain the cis-acting nucleolus organizer regions responsible for forming the nucleolus and the genes for the 28S, 18S, and 5.8S/2S RNA components of the ribosomes and so serve a central role in protein synthesis. Mutations or alterations that affect the nucleolus organizer region have pleiotropic effects on genome regulation and development and may play a role in genomewide phenomena such as aging and cancer. We demonstrate a method to create an allelic series of graded deletions in the Drosophila Y-linked rDNA of otherwise isogenic chromosomes, quantify the size of the deletions using real-time PCR, and monitor magnification of the rDNA arrays as their functions are restored. We use this series to define the thresholds of Y-linked rDNA required for sufficient protein translation, as well as establish the rate of Y-linked rDNA magnification in Drosophila. Finally, we show that I-CreI expression can revert rDNA deletion phenotypes, suggesting that double-strand breaks are sufficient to induce rDNA magnification.

Original languageEnglish (US)
Pages (from-to)1661-1671
Number of pages11
JournalGenetics
Volume181
Issue number4
DOIs
StatePublished - Apr 2009
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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