RT Journal
A1 Piganeau, Marion
A1 Ghezraoui, Hind
A1 De Cian, Anne
A1 Guittat, Lionel
A1 Tomishima, Mark
A1 Perrouault, Loic
A1 René, Oliver
A1 Katibah, George E.
A1 Zhang, Lei
A1 Holmes, Michael C.
A1 Doyon, Yannick
A1 Concordet, Jean-Paul
A1 Giovannangeli, Carine
A1 Jasin, Maria
A1 Brunet, Erika
T1 Cancer translocations in human cells induced by zinc finger and TALE nucleases
JF Genome Research 
JO Genome Research 
YR 2013 
FD July 01 
VO 23 
IS 7 
SP 1182 
OP 1193 
DO 10.1101/gr.147314.112 
UL http://genome.cshlp.org/content/23/7/1182.abstract 
AB Chromosomal translocations are signatures of numerous cancers and lead to expression of fusion genes that act as oncogenes. The wealth of genomic aberrations found in cancer, however, makes it challenging to assign a specific phenotypic change to a specific aberration. In this study, we set out to use genome editing with zinc finger (ZFN) and transcription activator-like effector (TALEN) nucleases to engineer, de novo, translocation-associated oncogenes at cognate endogenous loci in human cells. Using ZFNs and TALENs designed to cut precisely at relevant translocation breakpoints, we induced cancer-relevant t(11;22)(q24;q12) and t(2;5)(p23;q35) translocations found in Ewing sarcoma and anaplastic large cell lymphoma (ALCL), respectively. We recovered both translocations with high efficiency, resulting in the expression of the EWSR1–FLI1 and NPM1–ALK fusions. Breakpoint junctions recovered after ZFN cleavage in human embryonic stem (ES) cell–derived mesenchymal precursor cells fully recapitulated the genomic characteristics found in tumor cells from Ewing sarcoma patients. This approach with tailored nucleases demonstrates that expression of fusion genes found in cancer cells can be induced from the native promoter, allowing interrogation of both the underlying mechanisms and oncogenic consequences of tumor-related translocations in human cells. With an analogous strategy, the ALCL translocation was reverted in a patient cell line to restore the integrity of the two participating chromosomes, further expanding the repertoire of genomic rearrangements that can be engineered by tailored nucleases.