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Improved recombination efficiency by inhibition of NHEJ DNA repair.

Schutzrecht WO 2012/168307 A2 (2012)
Verlagsversion Volltext
The present invention relates to a method for modifying a target sequence in the genome of a mammalian cell, the method comprising the step of introducing into a mammalian cell: a. one or more compounds that introduce double-strand breaks in said target sequence; b. one or more DNA molecules comprising a donor DNA sequence to be incorporated by homologous recombination into the genomic DNA of said mammalian cell within said target sequence, wherein said donor DNA sequence is flanked upstream by a first flanking element and downstream by a second flanking element, wherein said first and second flanking element are different and wherein each of said first and second flanking sequence are homologous to a continuous DNA sequence on either side of the double-strand break introduced by said one or more compounds of a. within said target sequence in the genome of said mammalian cell; and c. one or more compounds that decrease the activity of the non-homologous end joining (NHEJ) DNA repair complex in said mammalian cell. Further, the invention relates to a method of producing a non-human mammal carrying a modified target sequence in its genome.
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Publikationstyp Sonstiges: Patent
Veröffentlichungsdatum 2012-12-13
Veröffentlichungsnummer WO 2012/168307 A2
Anmeldedatum 2012-06-06
Anmelder/Inhaber Kühn, Ralf; Wurst, W.; Helmholtz Zentrum München, Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH)
Anmeldeland EP
Priorität EP11004637