Daboussi, Fayza, Zaslavskiy, Mikhail, Poirot, Laurent, Loperfido, Mariana, Gouble, Agnès, Guyot, Valerie, Leduc, Sophie, Galetto, Roman, Grizot, Sylvestre, Oficjalska, Danusia, Perez, Christophe, Delacôte, Fabien, Dupuy, Aurélie, Chion-Sotinel, Isabelle, Le Clerre, Diane, Lebuhotel, Céline, Danos, Olivier, Lemaire, Frédéric, Oussedik, Kahina, Cédrone, Frédéric, Epinat, Jean-Charles, Smith, Julianne, Dickson, George, Popplewell, Linda, Koo, Taeyoung, Vandendriessche, Thierry, Chuah, Marinee K, Duclert, Aymeric, Duchateau, Philippe and Pâques, Frédéric (2012) Chromosomal context and epigenetic mechanisms control the efficacy of genome editing by rare-cutting designer endonucleases. Nucleic Acids Research
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The ability to specifically engineer the genome of living cells at precise locations using rare-cutting designer endonucleases has broad implications for biotechnology and medicine, particularly for functional genomics, transgenics and gene therapy. However, the potential impact of chromosomal context and epigenetics on designer endonuclease-mediated genome editing is poorly understood. To address this question, we conducted a comprehensive analysis on the efficacy of 37 endonucleases derived from the quintessential I-CreI meganuclease that were specifically designed to cleave 39 different genomic targets. The analysis revealed that the efficiency of targeted mutagenesis at a given chromosomal locus is predictive of that of homologous gene targeting. Consequently, a strong genome-wide correlation was apparent between the efficiency of targeted mutagenesis (≤0.1% to ∼6%) with that of homologous gene targeting (≤0.1% to ∼15%). In contrast, the efficiency of targeted mutagenesis or homologous gene targeting at a given chromosomal locus does not correlate with the activity of individual endonucleases on transiently transfected substrates. Finally, we demonstrate that chromatin accessibility modulates the efficacy of rare-cutting endonucleases, accounting for strong position effects. Thus, chromosomal context and epigenetic mechanisms may play a major role in the efficiency rare-cutting endonuclease-induced genome engineering.
This is a Submitted version This version's date is: 2012 This item is not peer reviewed
https://repository.royalholloway.ac.uk/items/058505dc-7d01-2380-fdd3-7ceaabbcb4f2/1/
Deposited by Research Information System (atira) on 24-May-2012 in Royal Holloway Research Online.Last modified on 24-May-2012