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Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the “safe harbor” AAVS1 locus
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 rema...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5446316/ https://www.ncbi.nlm.nih.gov/pubmed/28677529 http://dx.doi.org/10.1016/j.scr.2017.04.011 |
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author | Castaño, Julio Bueno, Clara Jiménez-Delgado, Senda Roca-Ho, Heleia Fraga, Mario F. Fernandez, Agustín F. Nakanishi, Mahito Torres-Ruiz, Raúl Rodríguez-Perales, Sandra Menéndez, Pablo |
author_facet | Castaño, Julio Bueno, Clara Jiménez-Delgado, Senda Roca-Ho, Heleia Fraga, Mario F. Fernandez, Agustín F. Nakanishi, Mahito Torres-Ruiz, Raúl Rodríguez-Perales, Sandra Menéndez, Pablo |
author_sort | Castaño, Julio |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5446316 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-54463162017-05-31 Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the “safe harbor” AAVS1 locus Castaño, Julio Bueno, Clara Jiménez-Delgado, Senda Roca-Ho, Heleia Fraga, Mario F. Fernandez, Agustín F. Nakanishi, Mahito Torres-Ruiz, Raúl Rodríguez-Perales, Sandra Menéndez, Pablo Stem Cell Res Lab Resource: Stem Cell Line 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. Elsevier 2017-05 /pmc/articles/PMC5446316/ /pubmed/28677529 http://dx.doi.org/10.1016/j.scr.2017.04.011 Text en © 2017 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Lab Resource: Stem Cell Line Castaño, Julio Bueno, Clara Jiménez-Delgado, Senda Roca-Ho, Heleia Fraga, Mario F. Fernandez, Agustín F. Nakanishi, Mahito Torres-Ruiz, Raúl Rodríguez-Perales, Sandra Menéndez, Pablo Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the “safe harbor” AAVS1 locus |
title | Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the “safe harbor” AAVS1 locus |
title_full | Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the “safe harbor” AAVS1 locus |
title_fullStr | Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the “safe harbor” AAVS1 locus |
title_full_unstemmed | Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the “safe harbor” AAVS1 locus |
title_short | Generation and characterization of a human iPSC cell line expressing inducible Cas9 in the “safe harbor” AAVS1 locus |
title_sort | generation and characterization of a human ipsc cell line expressing inducible cas9 in the “safe harbor” aavs1 locus |
topic | Lab Resource: Stem Cell Line |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5446316/ https://www.ncbi.nlm.nih.gov/pubmed/28677529 http://dx.doi.org/10.1016/j.scr.2017.04.011 |
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