Simple and Rapid InVivo Generation of Chromosomal Rearrangements using CRISPR/Cas9 Technology
Articolo
Data di Pubblicazione:
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.
Tipologia CRIS:
03A-Articolo su Rivista
Keywords:
Animals; Base Sequence; CRISPR-Associated Proteins; Carcinogenesis; Clustered Regularly Interspaced Short Palindromic Repeats; Genetic Engineering; HEK293 Cells; Humans; Lung Neoplasms; Mice; Molecular Sequence Data; Oncogene Proteins, Fusion; RNA, Messenger; Translocation, Genetic; Gene Rearrangement; Biochemistry, Genetics and Molecular Biology (all); Medicine (all)
Elenco autori:
Blasco, Rafael B; Karaca, Elif; Ambrogio, Chiara; Cheong, Taek-Chin; Karayol, Emre; Minero, Valerio G.; Voena, Claudia; Chiarle, Roberto
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