Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 Technology
Recent advances in genome engineering based on the CRISPR/Cas9 technology have revolutionized our ability to manipulate genomic DNA. Its use in human pluripotent stem cells (hPSCs) has allowed a wide range of mutant cell lines to be obtained at an unprecedented rate. The combination of these two gro...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-05-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/9/5/1312 |
| _version_ | 1797567109298913280 |
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| author | Joffrey Mianné Chloé Bourguignon Chloé Nguyen Van Mathieu Fieldès Amel Nasri Said Assou John De Vos |
| author_facet | Joffrey Mianné Chloé Bourguignon Chloé Nguyen Van Mathieu Fieldès Amel Nasri Said Assou John De Vos |
| author_sort | Joffrey Mianné |
| collection | DOAJ |
| description | Recent advances in genome engineering based on the CRISPR/Cas9 technology have revolutionized our ability to manipulate genomic DNA. Its use in human pluripotent stem cells (hPSCs) has allowed a wide range of mutant cell lines to be obtained at an unprecedented rate. The combination of these two groundbreaking technologies has tremendous potential, from disease modeling to stem cell-based therapies. However, the generation, screening and molecular characterization of these cell lines remain a cumbersome and multi-step endeavor. Here, we propose a pipeline of strategies to efficiently generate, sub-clone, and characterize CRISPR/Cas9-edited hPSC lines in the function of the introduced mutation (indels, point mutations, insertion of large constructs, deletions). |
| first_indexed | 2024-03-10T19:36:59Z |
| format | Article |
| id | doaj.art-cd214da837c84b0581f234ac03faec96 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T19:36:59Z |
| publishDate | 2020-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-cd214da837c84b0581f234ac03faec962023-11-20T01:36:56ZengMDPI AGCells2073-44092020-05-0195131210.3390/cells9051312Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 TechnologyJoffrey Mianné0Chloé Bourguignon1Chloé Nguyen Van2Mathieu Fieldès3Amel Nasri4Said Assou5John De Vos6IRMB, Univ Montpellier, INSERM, CHU Montpellier, 34000 Montpellier, FranceIRMB, Univ Montpellier, INSERM, CHU Montpellier, 34000 Montpellier, FranceIRMB, Univ Montpellier, INSERM, CHU Montpellier, 34000 Montpellier, FranceIRMB, Univ Montpellier, INSERM, CHU Montpellier, 34000 Montpellier, FranceIRMB, Univ Montpellier, INSERM, CHU Montpellier, 34000 Montpellier, FranceIRMB, Univ Montpellier, INSERM, CHU Montpellier, 34000 Montpellier, FranceIRMB, Univ Montpellier, INSERM, CHU Montpellier, 34000 Montpellier, FranceRecent advances in genome engineering based on the CRISPR/Cas9 technology have revolutionized our ability to manipulate genomic DNA. Its use in human pluripotent stem cells (hPSCs) has allowed a wide range of mutant cell lines to be obtained at an unprecedented rate. The combination of these two groundbreaking technologies has tremendous potential, from disease modeling to stem cell-based therapies. However, the generation, screening and molecular characterization of these cell lines remain a cumbersome and multi-step endeavor. Here, we propose a pipeline of strategies to efficiently generate, sub-clone, and characterize CRISPR/Cas9-edited hPSC lines in the function of the introduced mutation (indels, point mutations, insertion of large constructs, deletions).https://www.mdpi.com/2073-4409/9/5/1312hPSCCRISPRscreeningstem cellsgene editinggene targeting |
| spellingShingle | Joffrey Mianné Chloé Bourguignon Chloé Nguyen Van Mathieu Fieldès Amel Nasri Said Assou John De Vos Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 Technology Cells hPSC CRISPR screening stem cells gene editing gene targeting |
| title | Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 Technology |
| title_full | Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 Technology |
| title_fullStr | Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 Technology |
| title_full_unstemmed | Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 Technology |
| title_short | Pipeline for the Generation and Characterization of Transgenic Human Pluripotent Stem Cells Using the CRISPR/Cas9 Technology |
| title_sort | pipeline for the generation and characterization of transgenic human pluripotent stem cells using the crispr cas9 technology |
| topic | hPSC CRISPR screening stem cells gene editing gene targeting |
| url | https://www.mdpi.com/2073-4409/9/5/1312 |
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