Développement de nouvelles approches d’édition du génome à l’aide de nucléases artificielles (TALENs et CRISPR/Cas9)

Abstract : Genome editing relies on the ability of artificial nucleases (TALEN or CRISPR/Cas9 system) to induce double strand break into a precise and unique sequence in a whole genome and on the different DNA repair system. The two major DNA repair systems are NHEJ (Non Homologous End Joining) and HR (Homologous Recombination). NHEJ consists on DNA end direct ligation. This system can lead to deletion or insertion at the cut site. These mutations, when induced in an exon, can induce reading frame change and gene inactivation (Knock out). HR consists on the use of sister chromatid to copy lost information in order to complete the double strand break. If an exogenous DNA with homologies with the targeted DNA is inserted with artificial nucleases, it can be used as a template and can permit to introduce any transgene at the cut site (Knock In). In this work, different strategies were used to optimize genome editing. By fusing Nter part of CtIP to Cas9, the KI rate of an exogenous DNA is increased and by fusing Trex2 exonuclease to Cas9, the mutation rate induced is also increased. These two approaches can be widely used to improve genome editing strategies.
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Marine Charpentier. Développement de nouvelles approches d’édition du génome à l’aide de nucléases artificielles (TALENs et CRISPR/Cas9). Génétique. École pratique des hautes études - EPHE PARIS, 2016. Français. ⟨NNT : 2016EPHE3106⟩. ⟨tel-02105233⟩

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