title: Simple and rapid in vivo generation of chromosomal rearrangements using CRISPR/Cas9 technology.
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.
authors.affiliation: Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Rafael B
authors.initials: RB
authors.lastname: Blasco
authors.affiliation: Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Elif
authors.initials: E
authors.lastname: Karaca
authors.affiliation: Molecular Oncology Program, Centro Nacional de Investigaciones Oncológicas, 28029 Madrid, Spain.
authors.forename: Chiara
authors.initials: C
authors.lastname: Ambrogio
authors.affiliation: Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Taek-Chin
authors.initials: TC
authors.lastname: Cheong
authors.affiliation: Department of Pathology, Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Emre
authors.initials: E
authors.lastname: Karayol
authors.affiliation: Department of Molecular Biotechnology and Health Sciences, University of Torino, 10126 Torino, Italy.
authors.forename: Valerio G
authors.initials: VG
authors.lastname: Minero
authors.affiliation: 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.
authors.forename: Claudia
authors.initials: C
authors.lastname: Voena
authors.affiliation: 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.
authors.forename: Roberto
authors.initials: R
authors.lastname: Chiarle
mesh_terms: D000818:Animals; D001483:Base Sequence; D064130:CRISPR-Associated Proteins / Q000378:metabolism*; D063646:Carcinogenesis / Q000235:genetics / Q000473:pathology; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D015321:Gene Rearrangement; D005818:Genetic Engineering / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D015514:Oncogene Proteins, Fusion / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014178:Translocation, Genetic / Q000235:genetics*
chemical_list: D064130:CRISPR-Associated Proteins; D015514:Oncogene Proteins, Fusion; D012333:RNA, Messenger