HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug Control
The CRISPR/Cas9 enabled efficient gene editing in an easy and programmable manner. Controlling its activity in greater precision is desired for biomedical research and potential therapeutic translation. Here, we engrafted the CRISPR/Cas9 system with a mutated human estrogen receptor (ERT2), which re...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2018-12-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253118302373 |
| _version_ | 1819144492094062592 |
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| author | Chen Zhao Yingze Zhao Jingfang Zhang Jia Lu Li Chen Yue Zhang Yue Ying Junjun Xu Shixian Wei Yu Wang |
| author_facet | Chen Zhao Yingze Zhao Jingfang Zhang Jia Lu Li Chen Yue Zhang Yue Ying Junjun Xu Shixian Wei Yu Wang |
| author_sort | Chen Zhao |
| collection | DOAJ |
| description | The CRISPR/Cas9 enabled efficient gene editing in an easy and programmable manner. Controlling its activity in greater precision is desired for biomedical research and potential therapeutic translation. Here, we engrafted the CRISPR/Cas9 system with a mutated human estrogen receptor (ERT2), which renders it 4-hydroxytamoxifen (4-OHT) inducible for the access of genome, and a nuclear export signal (NES), which lowers the background activity. Tight and efficient drug-inducible genome editing was achieved across several human cell types, including embryonic stem cells (ESCs) and mesenchymal stem cells (MSCs), upon vigorous optimization. Optimized terminal device, which we named hybrid drug inducible CRISPR/Cas9 technology (HIT-Cas9), delivered advantageous performances over several existing designs. Such architecture was also successfully applied to an orthogonal Cas9. The HIT-Cas9 system developed in this study will find broad utility in controlled editing of potentially any genomic loci. Keywords: CRISPR/Cas9, genome editing, drug inducible, stem cells |
| first_indexed | 2024-12-22T12:42:59Z |
| format | Article |
| id | doaj.art-83c17ed66aec462cbae682d6f182737b |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-22T12:42:59Z |
| publishDate | 2018-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-83c17ed66aec462cbae682d6f182737b2022-12-21T18:25:23ZengElsevierMolecular Therapy: Nucleic Acids2162-25312018-12-0113208219HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug ControlChen Zhao0Yingze Zhao1Jingfang Zhang2Jia Lu3Li Chen4Yue Zhang5Yue Ying6Junjun Xu7Shixian Wei8Yu Wang9State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, ChinaState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, ChinaState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, ChinaState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, ChinaState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, ChinaState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, ChinaState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, ChinaCollege of Biological Sciences, China Agricultural University, Beijing 100193, ChinaState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, ChinaState Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China; Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China; Corresponding author: Yu Wang, State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.The CRISPR/Cas9 enabled efficient gene editing in an easy and programmable manner. Controlling its activity in greater precision is desired for biomedical research and potential therapeutic translation. Here, we engrafted the CRISPR/Cas9 system with a mutated human estrogen receptor (ERT2), which renders it 4-hydroxytamoxifen (4-OHT) inducible for the access of genome, and a nuclear export signal (NES), which lowers the background activity. Tight and efficient drug-inducible genome editing was achieved across several human cell types, including embryonic stem cells (ESCs) and mesenchymal stem cells (MSCs), upon vigorous optimization. Optimized terminal device, which we named hybrid drug inducible CRISPR/Cas9 technology (HIT-Cas9), delivered advantageous performances over several existing designs. Such architecture was also successfully applied to an orthogonal Cas9. The HIT-Cas9 system developed in this study will find broad utility in controlled editing of potentially any genomic loci. Keywords: CRISPR/Cas9, genome editing, drug inducible, stem cellshttp://www.sciencedirect.com/science/article/pii/S2162253118302373 |
| spellingShingle | Chen Zhao Yingze Zhao Jingfang Zhang Jia Lu Li Chen Yue Zhang Yue Ying Junjun Xu Shixian Wei Yu Wang HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug Control Molecular Therapy: Nucleic Acids |
| title | HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug Control |
| title_full | HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug Control |
| title_fullStr | HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug Control |
| title_full_unstemmed | HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug Control |
| title_short | HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug Control |
| title_sort | hit cas9 a crispr cas9 genome editing device under tight and effective drug control |
| url | http://www.sciencedirect.com/science/article/pii/S2162253118302373 |
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