Generation of chromosomal translocations that lead to conditional fusion protein expression using CRISPR-Cas9 and homology-directed repair Journal Article


Authors: Vanoli, F.; Jasin, M.
Article Title: Generation of chromosomal translocations that lead to conditional fusion protein expression using CRISPR-Cas9 and homology-directed repair
Abstract: Recurrent chromosomal translocations often lead to expression of fusion proteins associated with oncogenic transformation. To study translocations and downstream events, genome editing techniques have been developed to generate chromosomal translocations through non-homologous end joining of DNA double-strand breaks introduced at the two participating endogenous loci. However, the frequencies at which these events occur is usually too low to efficiently clone cells carrying the translocation. This article provides a detailed method using CRISPR-Cas9 technology and homology-directed repair to efficiently isolate cells harboring a chromosomal translocation. For an additional level of control, the resulting fusion protein is conditionally expressed to allow early events in oncogenic transformation to be studied. We focus on the generation of the EWSR1-WT1 fusion using human mesenchymal cells, which is associated with the translocation found in desmoplastic small round cell tumors. © 2017 Elsevier Inc.
Keywords: double-strand break; chromosomal translocation; hdr; crispr-cas9; ewsr1-wt1
Journal Title: Methods
Volume: 121-122
ISSN: 1046-2023
Publisher: Academic Press Inc., Elsevier Science  
Date Published: 2017-05-15
Start Page: 138
End Page: 145
Language: English
DOI: 10.1016/j.ymeth.2017.05.006
PROVIDER: scopus
PMCID: PMC5531069
PUBMED: 28522325
DOI/URL:
Notes: Article -- Export Date: 1 August 2017 -- Source: Scopus
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MSK Authors
  1. Maria Jasin
    213 Jasin
  2. Fabio Vanoli
    8 Vanoli