Engineering human tumour-associated chromosomal translocations with the RNA-guided CRISPR–Cas9 system

R. Torres, M. C. Martin, A. Garcia, Juan C. Cigudosa, J. C. Ramirez () and S. Rodriguez-Perales ()
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R. Torres: Viral Vector Facility, Fundacion Centro Nacional de Investigaciones Cardiovasculares (CNIC), Melchor Fernandez Almagro 3
M. C. Martin: Molecular Cytogenetics Group, Spanish National Cancer Centre-CNIO, Melchor Fernandez Almagro 3
A. Garcia: Viral Vector Facility, Fundacion Centro Nacional de Investigaciones Cardiovasculares (CNIC), Melchor Fernandez Almagro 3
Juan C. Cigudosa: Molecular Cytogenetics Group, Spanish National Cancer Centre-CNIO, Melchor Fernandez Almagro 3
J. C. Ramirez: Viral Vector Facility, Fundacion Centro Nacional de Investigaciones Cardiovasculares (CNIC), Melchor Fernandez Almagro 3
S. Rodriguez-Perales: Molecular Cytogenetics Group, Spanish National Cancer Centre-CNIO, Melchor Fernandez Almagro 3

Nature Communications, 2014, vol. 5, issue 1, 1-8

Abstract: Abstract Cancer-related human chromosomal translocations are generated through the illegitimate joining of two non-homologous chromosomes affected by double-strand breaks (DSB). Effective methodologies to reproduce precise reciprocal tumour-associated chromosomal translocations are required to gain insight into the initiation of leukaemia and sarcomas. Here we present a strategy for generating cancer-related human chromosomal translocations in vitro based on the ability of the RNA-guided CRISPR–Cas9 system to induce DSBs at defined positions. Using this approach we generate human cell lines and primary cells bearing chromosomal translocations resembling those described in acute myeloid leukaemia and Ewing’s sarcoma at high frequencies. FISH and molecular analysis at the mRNA and protein levels of the fusion genes involved in these engineered cells reveal the reliability and accuracy of the CRISPR–Cas9 approach, providing a powerful tool for cancer studies.

Date: 2014
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DOI: 10.1038/ncomms4964

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