Chromosomal context and epigenetic mechanisms control the efficacy of genome editing by rare-cutting designer endonucleases

Fayza Daboussi, Mikhail Zaslavskiy, Laurent Poirot, Mariana Loperfido, Agnès Gouble, Valerie Guyot, Sophie Leduc, Roman Galetto, Sylvestre Grizot, Danusia Oficjalska, Christophe Perez, Fabien Delacôte, Aurélie Dupuy, Isabelle Chion-Sotinel, Diane Le Clerre, Céline Lebuhotel, Olivier Danos, Frédéric Lemaire, Kahina Oussedik, Frédéric CédroneJean-Charles Epinat, Julianne Smith, Rafael J Yáñez-Muñoz, George Dickson, Linda Popplewell, Taeyoung Koo, Thierry VandenDriessche, Marinee K Chuah, Aymeric Duclert, Philippe Duchateau, Frédéric Pâques

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)6367–6379
Number of pages13
JournalNucleic Acids Research
Volume40
Issue number13
Early online date28 Mar 2012
DOIs
Publication statusPublished - 1 Jul 2012
Externally publishedYes

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