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Title

Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the “safe harbor” AAVS1 locus

AuthorsCastaño, Julio; Bueno, Clara; Jiménez-Delgado, Senda; Roca-Ho, Heleia; Fraga, Mario F.; Fernández, Agustín F.; Nakanishi, Mahito; Torres-Ruiz, Raúl; Rodríguez-Perales, Sandra; Menéndez, Pablo
Issue Date2017
PublisherElsevier
CitationStem Cell Research 21: 137-140 (2017)
AbstractWe report the generation-characterization of a fetal liver (FL) B-cell progenitor (BCP)-derived human induced pluripotent stem cell (hiPSC) line CRISPR/Cas9-edited to carry/express a single copy of doxycycline-inducible Cas9 gene in the “safe locus” AAVS1 (iCas9-FL-BCP-hiPSC). Gene-edited iPSCs remained pluripotent after CRISPR/Cas9 genome-edition. Correct genomic integration of a unique copy of Cas9 was confirmed by PCR and Southern blot. Cas9 was robustly and specifically expressed on doxycycline exposure. T7-endonuclease assay demonstrated that iCas9 induces robust gene-edition when gRNAs against hematopoietic transcription factors were tested. This iCas9-FL-BCP-hiPSC will facilitate gene-editing approaches for studies on developmental biology, drug screening and disease modeling.
Publisher version (URL)https://doi.org/10.1016/j.scr.2017.04.011
URIhttp://hdl.handle.net/10261/173453
Identifiersdoi: 10.1016/j.scr.2017.04.011
e-issn: 1876-7753
issn: 1873-5061
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