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Updated: Mar 7, 2026

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

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Generation of conditional oncogenic chromosomal translocations using CRISPR-Cas9 genomic editing and

Lee Spraggon1, Luciano G Martelotto1, Julija Hmeljak1

  • 1Department of Pathology, Memorial Sloan Kettering Cancer Center, New York, NY, USA.

The Journal of Pathology
|February 12, 2017
PubMed

Insights

Researchers engineered chromosomal translocations using CRISPR-Cas9 technology to study cancer-driving fusion genes. This novel method allows temporal control over engineered rearrangements, aiding in the development of new cancer models.

Area of Science:

  • Molecular Biology
  • Genetics
  • Oncology

Background:

  • Chromosomal rearrangements creating oncogenic fusion proteins are hallmarks of many cancers.
  • Previous methods utilized programmable nucleases and DNA repair to generate translocations, leading to constitutively expressed fusion genes.

Purpose of the Study:

  • To develop a novel approach combining CRISPR-Cas9 and homology-directed repair for engineering chromosomal translocations.
  • To enable temporal control over the expression of engineered chromosomal rearrangements.
  • To create models for studying tumors driven by fusion genes.

Main Methods:

  • Utilized CRISPR-Cas9 technology combined with homology-directed repair.
  • Engineered, captured, and modulated expression of chromosomal translocation products in a human cell line.
  • Applied the approach to model translocations t(11;22)(q24;q12) and t(11;22)(p13;q12) involving EWSR1 gene fusions.

Main Results:

  • Successfully engineered chromosomal translocations in a human cell line.
  • Demonstrated temporal control over the expression of engineered endogenous chromosomal rearrangements.
  • Generated genetic models for specific translocations found in Ewing's sarcoma and desmoplastic small round cell tumor.

Conclusions:

  • The novel CRISPR-Cas9 based approach provides precise control over chromosomal translocation engineering.
  • This method facilitates the creation of advanced cellular models for studying fusion gene-driven malignancies.
  • The technique offers a powerful tool for investigating the mechanisms of tumorigenesis driven by chromosomal rearrangements.
Keywords:
CRISPR-Cas9Ewing sarcomachromosomal translocationsdesmoplastic small round cell tumour (DSRCT)genomic editinghomology-directed repaironcogene

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