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Simple and rapid in vivo generation of chromosomal rearrangements using CRISPR/Cas9 technology.
- 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.)
- Subjects :
- Animals
Base Sequence
Carcinogenesis genetics
Carcinogenesis pathology
HEK293 Cells
Humans
Lung Neoplasms genetics
Mice
Molecular Sequence Data
Oncogene Proteins, Fusion genetics
Oncogene Proteins, Fusion metabolism
RNA, Messenger genetics
RNA, Messenger metabolism
CRISPR-Associated Proteins metabolism
Clustered Regularly Interspaced Short Palindromic Repeats genetics
Gene Rearrangement
Genetic Engineering methods
Translocation, Genetic genetics
Subjects
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