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Cancer translocations in human cells induced by zinc finger and TALE nucleases

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 u...

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Autores principales: Piganeau, Marion, Ghezraoui, Hind, De Cian, Anne, Guittat, Lionel, Tomishima, Mark, Perrouault, Loic, René, Oliver, Katibah, George E., Zhang, Lei, Holmes, Michael C., Doyon, Yannick, Concordet, Jean-Paul, Giovannangeli, Carine, Jasin, Maria, Brunet, Erika
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3698511/
https://www.ncbi.nlm.nih.gov/pubmed/23568838
http://dx.doi.org/10.1101/gr.147314.112
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author Piganeau, Marion
Ghezraoui, Hind
De Cian, Anne
Guittat, Lionel
Tomishima, Mark
Perrouault, Loic
René, Oliver
Katibah, George E.
Zhang, Lei
Holmes, Michael C.
Doyon, Yannick
Concordet, Jean-Paul
Giovannangeli, Carine
Jasin, Maria
Brunet, Erika
author_facet Piganeau, Marion
Ghezraoui, Hind
De Cian, Anne
Guittat, Lionel
Tomishima, Mark
Perrouault, Loic
René, Oliver
Katibah, George E.
Zhang, Lei
Holmes, Michael C.
Doyon, Yannick
Concordet, Jean-Paul
Giovannangeli, Carine
Jasin, Maria
Brunet, Erika
author_sort Piganeau, Marion
collection PubMed
description 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.
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spelling pubmed-36985112013-07-03 Cancer translocations in human cells induced by zinc finger and TALE nucleases Piganeau, Marion Ghezraoui, Hind De Cian, Anne Guittat, Lionel Tomishima, Mark Perrouault, Loic René, Oliver Katibah, George E. Zhang, Lei Holmes, Michael C. Doyon, Yannick Concordet, Jean-Paul Giovannangeli, Carine Jasin, Maria Brunet, Erika Genome Res Method 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. Cold Spring Harbor Laboratory Press 2013-07 /pmc/articles/PMC3698511/ /pubmed/23568838 http://dx.doi.org/10.1101/gr.147314.112 Text en © 2013, Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/3.0/ This article, published in Genome Research, is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.
spellingShingle Method
Piganeau, Marion
Ghezraoui, Hind
De Cian, Anne
Guittat, Lionel
Tomishima, Mark
Perrouault, Loic
René, Oliver
Katibah, George E.
Zhang, Lei
Holmes, Michael C.
Doyon, Yannick
Concordet, Jean-Paul
Giovannangeli, Carine
Jasin, Maria
Brunet, Erika
Cancer translocations in human cells induced by zinc finger and TALE nucleases
title Cancer translocations in human cells induced by zinc finger and TALE nucleases
title_full Cancer translocations in human cells induced by zinc finger and TALE nucleases
title_fullStr Cancer translocations in human cells induced by zinc finger and TALE nucleases
title_full_unstemmed Cancer translocations in human cells induced by zinc finger and TALE nucleases
title_short Cancer translocations in human cells induced by zinc finger and TALE nucleases
title_sort cancer translocations in human cells induced by zinc finger and tale nucleases
topic Method
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3698511/
https://www.ncbi.nlm.nih.gov/pubmed/23568838
http://dx.doi.org/10.1101/gr.147314.112
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