Controlled Activation of LINE-1 Retrotransposons in Mammals
LINE-1 (Long INterspersed Element-1) elements represent the major class of mammalian non-LTR (long terminal repeat) retrotransposons which are cov-ering 35 % of the human genome. LINE-1 is the only autonomous non-LTR retrotransposon in the human genome because it is coding for the protein machinery required for its mobilization. <br><br>
We offer a system which allows inducible, controlled and conditional expres-sion and retrotransposition of non-LTR retrotransposons, in particular mam-malian LINE-1 elements, in mammalian systems (cell culture and in vivo). The LINE-1 element is tagged with a reporter gene cassette and set under control of a tetracycline/doxycycline-inducible promoter. The modified LINE-1 elements can be used for an efficient insertional mutagenesis screen in both germ line and somatic cells. Gene trapping approaches based on controlled retrotransposition during defined time periods of the embryonic development can be used to identify cancer genes in specific tissues and cell types. Inducible modified LINE-1 elements which are tagged with a therapeutic gene can be used as a gene delivery system in gene therapeutic approaches.<br><br>
<li>Induction of L1 retrotransposition for a defined period of time</li>
<li>Tissue-specific L1 retrotransposition induction possible by using
a vector-construct, coding for rtTA or tTA functionally linked to a tissue-specific promoter
<li>Proof of principle Tet/Dox – On/Off system shown in M2-HeLa cells</li></ul>
Patent granted in DE, GB, FR (EP 2205750), US patent pending (priority date 10/2007).
Paul-Ehrlich-Institut, Federal Agency for Sera and Vaccines, Langen
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