Please use this identifier to cite or link to this item:http://hdl.handle.net/20.500.12105/7430
Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the "safe harbor" AAVS1 locus
Stem Cell Res. 2017;21:137-140
We 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.
B-Lymphocytes | Bacterial Proteins | CRISPR-Associated Protein 9 | CRISPR-Cas Systems | Endonucleases | Gene Editing | Humans | Induced Pluripotent Stem Cells | Dependovirus | Gene Expression | Genetic Loci
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