Analysis of intrachromosomal homologous recombination in mammalian cell, using tandem repeat sequences
Abstract
In all the organisms, homologous recombination HR is involved in fundamental processes such as genome diversification and DNA repair. Several strategies can be devised to measure homologous recombination in mammalian cells. We present here the interest of using intrachromosomal tandem repeat sequences to measure HR in mammalian cells and we discuss the differences with the ectopic plasmids recombination. The present review focuses on the molecular mechanisms of HR between tandem repeats in mammalian cells. The possibility to use two different orientations of tandem repeats direct or. inverted repeats in parallel constitutes also an advantage. While inverted repeats measure only events arising by strand exchange gene conversion and crossing over , direct repeats monitor strand exchange events and also non-conservative processes such as single strand annealing or replication slippage. In yeast, these processes depend on different pathways, most of them also existing in mammalian cells. These data permit to devise substrates adapted to specific questions about HR in mammalian cells. The effect of substrate structures heterologies, insertionsrdeletions, GT repeats, transcription and consequences of DNA double strand breaks induced by ionizing radiation or endonuclease especially the rare-cutting. endonuclease ISce-I on HR are discussed. Finally, transgenic mouse models using tandem repeats are briefly presented.
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