Generation of conditional oncogenic chromosomal translocations using CRISPR-Cas9 genomic editing and homology-directed repair. Academic Article uri icon

Overview

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.

publication date

  • March 30, 2017

Research

keywords

  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Desmoplastic Small Round Cell Tumor
  • Recombinational DNA Repair
  • Sarcoma, Ewing
  • Translocation, Genetic

Identity

PubMed Central ID

  • PMC5397343

Scopus Document Identifier

  • 85016967426

Digital Object Identifier (DOI)

  • 10.1002/path.4883

PubMed ID

  • 28188619

Additional Document Info

volume

  • 242

issue

  • 1