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| Subject Categories: Genome Stability & Dynamics | Genomic & Computational Biology |
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| The EMBO Journal
(2007) 26, 170–183, doi:10.1038/sj.emboj.7601463 Published online 7 December 2006 |
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| Targeted engineering of the Caenorhabditis elegans genome following Mos1-triggered chromosomal breaks |
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| Valérie Robert and Jean-Louis Bessereau |
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ENS, Biologie cellulaire de la synapse, Paris, France; Inserm, U789, Paris, France To whom correspondence should be addressed Jean-Louis Bessereau, Ecole Normale Supérieure, INSERM U789, 46 Rue d'Ulm, Paris 75005, France. Tel.: +33 1 44 32 23 05; Fax: +33 1 44 32 36 54; E-mail: jlbesse@biologie.ens.fr Received 19 June 2006; Accepted 2 November 2006; Published online 7 December 2006. |
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| Abstract |
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| The Drosophila element Mos1 is a class II transposon, which moves by a 'cut-and-paste' mechanism and can be experimentally mobilized in the Caenorhabditis elegans germ line. Here, we triggered the excision of identified Mos1 insertions to create chromosomal breaks at given sites and further manipulate the broken loci. Double-strand break (DSB) repair could be achieved by gene conversion using a transgene containing sequences homologous to the broken chromosomal region as a repair template. Consequently, mutations engineered in the transgene could be copied to a specific locus at high frequency. This pathway was further characterized to develop an efficient tool—called MosTIC—to manipulate the C. elegans genome. Analysis of DSB repair during MosTIC experiments demonstrated that DSBs could also be sealed by end-joining in the germ line, independently from the evolutionarily conserved Ku80 and ligase IV factors. In conjunction with a publicly available Mos1 insertion library currently being generated, MosTIC will provide a general tool to customize the C. elegans genome. |
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| Keywords: DSB repair, homologous recombination, NHEJ, targeted mutagenesis |
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