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Journal of Nucleic Acids
Volume 2011, Article ID 947212, 14 pages
Research Article

Identification of Genes Regulating Gene Targeting by a High-Throughput Screening Approach

1Cellectis SA, 102 Avenue Gaston Roussel, 93340 Romainville Cedex, France
2Cellectis Bioresearch, 102 Avenue Gaston Roussel, 93340 Romainville Cedex, France

Received 25 November 2010; Accepted 23 January 2011

Academic Editor: Emery H. Bresnick

Copyright © 2011 Fabien Delacôte et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Homologous gene targeting (HGT) is a precise but inefficient process for genome engineering. Several methods for increasing its efficiency have been developed, including the use of rare cutting endonucleases. However, there is still room for improvement, as even nuclease-induced HGT may vary in efficiency as a function of the nuclease, target site, and cell type considered. We have developed a high-throughput screening assay for the identification of factors stimulating meganuclease-induced HGT. We used this assay to explore a collection of siRNAs targeting 19,121 human genes. At the end of secondary screening, we had identified 64 genes for which knockdown affected nuclease-induced HGT. Two of the strongest candidates were characterized further. We showed that siRNAs directed against the ATF7IP gene, encoding a protein involved in chromatin remodeling, stimulated HGT by a factor of three to eight, at various loci and in different cell types. This method thus led to the identification of a number of genes, the manipulation of which might increase rates of targeted recombination.