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

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


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • CRISPR-Associated Proteins / metabolism*
  • Carcinogenesis / genetics
  • Carcinogenesis / pathology
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Gene Rearrangement*
  • Genetic Engineering / methods*
  • HEK293 Cells
  • Humans
  • Lung Neoplasms / genetics
  • Mice
  • Molecular Sequence Data
  • Oncogene Proteins, Fusion / genetics
  • Oncogene Proteins, Fusion / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Translocation, Genetic / genetics*


  • CRISPR-Associated Proteins
  • Oncogene Proteins, Fusion
  • RNA, Messenger