Generation of conditional oncogenic chromosomal translocations using CRISPR–Cas9 genomic editing and homology-directed repair Journal Article

   

Authors:	Spraggon, L.; Martelotto, L. G.; Hmeljak, J.; Hitchman, T. D.; Wang, J.; Wang, L.; Slotkin, E. K.; Fan, P. D.; Reis-Filho, J. S.; Ladanyi, M.
Article Title:	Generation of conditional oncogenic chromosomal translocations using CRISPR–Cas9 genomic editing and homology-directed repair
Abstract:	Chromosomal rearrangements encoding oncogenic fusion proteins are found in a wide variety of malignancies. The use of programmable nucleases to generate specific double-strand breaks in endogenous loci, followed by non-homologous end joining DNA repair, has allowed several of these translocations to be generated as constitutively expressed fusion genes within a cell population. Here, we describe a novel approach that combines CRISPR–Cas9 technology with homology-directed repair to engineer, capture, and modulate the expression of chromosomal translocation products in a human cell line. We have applied this approach to the genetic modelling of t(11;22)(q24;q12) and t(11;22)(p13;q12), translocation products of the EWSR1 gene and its 3' fusion partners FLI1 and WT1, present in Ewing's sarcoma and desmoplastic small round cell tumour, respectively. Our innovative approach allows for temporal control of the expression of engineered endogenous chromosomal rearrangements, and provides a means to generate models to study tumours driven by fusion genes. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Keywords:	ewing sarcoma; oncogene; chromosomal translocations; homology-directed repair; genomic editing; crispr–cas9; desmoplastic small round cell tumour (dsrct)
Journal Title:	Journal of Pathology
Volume:	242
Issue:	1
ISSN:	0022-3417
Publisher:	Wiley Blackwell
Date Published:	2017-05-01
Start Page:	102
End Page:	112
Language:	English
DOI:	10.1002/path.4883
PROVIDER:	scopus
PMCID:	PMC5397343
PUBMED:	28188619
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-85016967426&doi=10.1002%2fpath.4883&partnerID=40&md5=22e0f40e2f347d897dcc2b4932131771
Notes:	Article -- Export Date: 2 May 2017 -- Source: Scopus

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