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 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.
Würtele
Hugo
H
Institute of Research in Immunology and Cancer, Université de Montréal, Pavillon Roger-Gaudry, 2900 Edouard-Montpetit, Montreal, Quebec, Canada H3T 1J4.
Gusew
Nadine
N
Lussier
Roxane
R
Chartrand
Pierre
P
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004622:Embryo, Mammalian / Q000378:metabolism*; D005822:Genetic Vectors; D051379:Mice; D008822:Mice, Transgenic; D010957:Plasmids; D011995:Recombination, Genetic
D017931:DNA Primers