Characterization of in vivo recombination activities in the mouse embryo
Homologous recombination makes use of sequence homology to repair DNA and to rearrange genetic material. In mammals, these processes have mainly been characterized using cultured cell systems. We have developed an assay that allows us to quantitatively analyze homologous recombination in vivo in the...
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| Veröffentlicht in: | Molecular genetics and genomics : MGG 2005-05, Vol.273 (3), p.252-263 |
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| creator | Würtele, Hugo Gusew, Nadine Lussier, Roxane Chartrand, Pierre |
| description | Homologous recombination makes use of sequence homology to repair DNA and to rearrange genetic material. In mammals, these processes have mainly been characterized using cultured cell systems. We have developed an assay that allows us to quantitatively analyze homologous recombination in vivo in the mouse embryo. Transgenic mouse lines were generated by microinjection into a fertilized mouse ovum of a vector containing two homologous LINE-1 (L1) sequences arranged as a direct repeat: these sequences can recombine with each other and with endogenous L1 sequences before, during or after integration of the vector into the genome. Using a plasmid rescue procedure, we determined the composition of the integrated vector array in several transgenic mice and their descendants. Homologous recombination frequencies were found to be strikingly high, involving 70% of integrated vectors in some arrays, with homologous deletions being five times more frequent than gene conversion without crossing-over. Interestingly, non-homologous recombination was found to be much less frequent. We also found that endogenous L1 sequences could be involved in homologous recombination events in the mouse embryo, and that the integrated arrays could be modified from generation to generation by homologous recombination between the integrated L1 sequences. |
| doi_str_mv | 10.1007/s00438-005-1112-2 |
| format | Article |
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| subjects | Animals Base Sequence DNA Primers Embryo, Mammalian - metabolism Genetic Vectors Genetics Mice Mice, Transgenic Plasmids Recombination, Genetic Rodents |
| title | Characterization of in vivo recombination activities in the mouse embryo |
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