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Cell Rep. 2014 Nov 20;9(4):1219-27. doi: 10.1016/j.celrep.2014.10.051. Epub 2014 Nov 13.

Simple and rapid in vivo generation of chromosomal rearrangements using CRISPR/Cas9 technology.

Author information

1
Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
2
Molecular Oncology Program, Centro Nacional de Investigaciones Oncológicas, 28029 Madrid, Spain.
3
Department of Molecular Biotechnology and Health Sciences, University of Torino, 10126 Torino, Italy.
4
Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA; Department of Molecular Biotechnology and Health Sciences, University of Torino, 10126 Torino, Italy.
5
Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA; Department of Molecular Biotechnology and Health Sciences, University of Torino, 10126 Torino, Italy. Electronic address: roberto.chiarle@childrens.harvard.edu.

Abstract

Generation 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.

PMID:
25456124
DOI:
10.1016/j.celrep.2014.10.051
[Indexed for MEDLINE]
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