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

Authors :
Blasco RB
Karaca E
Ambrogio C
Cheong TC
Karayol E
Minero VG
Voena C
Chiarle R
Source :
Cell reports [Cell Rep] 2014 Nov 20; Vol. 9 (4), pp. 1219-27. Date of Electronic Publication: 2014 Nov 13.
Publication Year :
2014

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.<br /> (Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.)

Details

Language :
English
ISSN :
2211-1247
Volume :
9
Issue :
4
Database :
MEDLINE
Journal :
Cell reports
Publication Type :
Academic Journal
Accession number :
25456124
Full Text :
https://doi.org/10.1016/j.celrep.2014.10.051