2026-06-14T00:27:32Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/72382025-04-29T10:18:59Zcom_20.500.12105_2173com_20.500.12105_2051col_20.500.12105_19597
Repisalud
author
Torres-Ruiz Raul, Raul
author
Martinez-Lage, Marta
author
Martin, Maria C
author
Garcia, Aida
author
Bueno, Clara
author
Castaño, Julio
author
Ramirez, Juan C
author
Menéndez, Pablo
author
Cigudosa, Juan C
author
Rodriguez Perales, Sandra
funder
Instituto de Salud Carlos III
funder
Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF)
2019-02-26T12:56:37Z
2019-02-26T12:56:37Z
2017
Stem Cell Reports. 2017;8(5):1408-1420.
10.1016/j.stemcr.2017.04.014
2213-6711
22136711
Stem cell reports
28494941
http://hdl.handle.net/20.500.12105/7238
Efficient methodologies for recreating cancer-associated chromosome translocations are in high demand as tools for investigating how such events initiate cancer. The CRISPR/Cas9 system has been used to reconstruct the genetics of these complex rearrangements at native loci while maintaining the architecture and regulatory elements. However, the CRISPR system remains inefficient in human stem cells. Here, we compared three strategies aimed at enhancing the efficiency of the CRISPR-mediated t(11;22) translocation in human stem cells, including mesenchymal and induced pluripotent stem cells: (1) using end-joining DNA processing factors involved in repair mechanisms, or (2) ssODNs to guide the ligation of the double-strand break ends generated by CRISPR/Cas9; and (3) all-in-one plasmid or ribonucleoprotein complex-based approaches. We report that the generation of targeted t(11;22) is significantly increased by using a combination of ribonucleoprotein complexes and ssODNs. The CRISPR/Cas9-mediated generation of targeted t(11;22) in human stem cells opens up new avenues in modeling Ewing sarcoma.
eng
CRISPR
Cas9
Ewing sarcoma
MSC
Cancer modeling
Cancer translocation
Disease model
Genome engineering
Human stem cells
iPSC
Efficient Recreation of t(11;22) EWSR1-FLI1+ in Human Stem Cells Using CRISPR/Cas9
journal article
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