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Agenda de l'ENS de Lyon

KUCIAK - L1 retrotransposon activity : insights from genomic and molecular studies.

Soutenance de thèse

Jeudi 15 déc 2011



Description générale

L1 retrotransposons are the only autonomous and active transposable elements in humans and comprise as much as 20% of our DNA. They proliferate via an RNA intermediate and a coupled reverse transcription and integration process, called retrotransposition and mediated by an L1-encoded ribonucleoprotein particle (RNP). L1s are actively jumping in germ cells, embryonic stem cells and in the early embryo, occasionally leading to de novo genetic diseases, but are considered silent in most somatic tissues. To comprehensively map active L1 elements in the human genome and to further explore the importance and consequences of L1 retrotransposition in humans, we combined selective amplification of L1 insertion sites and high-throughput sequencing. We applied this strategy to obtain a differential map of L1 insertions in two related human cultured cell lines and to question the possibility that endogenous L1 elements could be jumping in somatic cultured cells. We discovered several L1 insertions only present in the daughter cell line but absent in the parental cell line, demonstrating for the first time that retrotransposition of endogenous L1s takes place in a human somatic cell line. To get insights into the determinants of L1 integration, we have also developed a novel reverse transcription assay using partially purified native L1 RNPs. This enabled us to show that the L1 reverse transcriptase participates to insertion site selection, adding a second layer of specificity beyond the L1 endonuclease. Finally our work highlights the flexibility of the L1 machinery, which certainly participates to the efficient spreading of L1 elements within mammalian genomes.


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