ArticleMethods (San Diego, Calif.)2017
Generation of chromosomal translocations that lead to conditional fusion protein expression using CRISPR-Cas9 and homology-directed repair.
Article in Methods (San Diego, Calif.), 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 17 citations in OpenAlex.
- Modeling Extraordinary Response Through Targeting Secondary Alterations in Fusion-Associated Sarcoma.JCO precision oncology · 2024Article
- Specificity of cancer-related chromosomal translocations is linked to proximity after the DNA double-strand break and subsequent selection.NAR cancer · 2023Article
- Review
- Developing Bottom-Up Induced Pluripotent Stem Cell Derived Solid Tumor Models Using Precision Genome Editing Technologies.The CRISPR journal · 2022Review
- Article
- Distinct pathways of homologous recombination controlled by the SWS1-SWSAP1-SPIDR complex.Nature communications · 2021Article
- Toward precise CRISPR DNA fragment editing and predictable 3D genome engineering.Journal of molecular cell biology · 2021Review
- The CRISP(Y) Future of Pediatric Soft Tissue Sarcomas.Frontiers in chemistry · 2020Review
- Translocation-Related Sarcomas.International journal of molecular sciences · 2018Review
- Precise Epigenome Editing on the Stage: A Novel Approach to Modulate Gene Expression.Epigenetics insights · 2018Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
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.
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Registered trials
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