• Rafael B Blasco, Elif Karaca, Chiara Ambrogio, Taek-Chin Cheong, Emre Karayol, Valerio G Minero, Claudia Voena, and Roberto Chiarle.
• Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
• Cell Rep. 2014 Nov 20; 9 (4): 1219-27.

AbstractGeneration of genetically engineered mouse models (GEMMs) for chromosomal translocations in the endogenous loci by a knockin strategy is lengthy and costly. The CRISPR/Cas9 system provides an innovative and flexible approach for genome engineering of genomic loci in vitro and in vivo. Here, we report the use of the CRISPR/Cas9 system for engineering a specific chromosomal translocation in adult mice in vivo. We designed CRISPR/Cas9 lentiviral vectors to induce cleavage of the murine endogenous Eml4 and Alk loci in order to generate the Eml4-Alk gene rearrangement recurrently found in non-small-cell lung cancers (NSCLCs). Intratracheal or intrapulmonary inoculation of lentiviruses induced Eml4-Alk gene rearrangement in lung cells in vivo. Genomic and mRNA sequencing confirmed the genome editing and the production of the Eml4-Alk fusion transcript. All mice developed Eml4-Alk-rearranged lung tumors 2 months after the inoculation, demonstrating that the CRISPR/Cas9 system is a feasible and simple method for the generation of chromosomal rearrangements in vivo.Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

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