Optical control of CRISPR-Cas editing with cyclically caged guide RNAs
ABSTRACT The CRISPR/Cas system has been proved as one of the most powerful tools for precise gene editing. However, the approaches for precise control over the genome editing and regulatory events are still desirable. Here, we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs. This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs. In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region. We have also achieved light-mediated MSTN gene editing in embryos, whereas a new bow-knot-type gRNA showed no background editing in the absence of light irradiation. Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited..
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Preprint| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
bioRxiv.org - (2022) vom: 20. Mai Zur Gesamtaufnahme - year:2022 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Sun, Ying-Jie [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.1101/2022.03.04.482981 |
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| funding: |
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XBI035428368 |
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| 245 | 1 | 0 | |a Optical control of CRISPR-Cas editing with cyclically caged guide RNAs |
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| 520 | |a ABSTRACT The CRISPR/Cas system has been proved as one of the most powerful tools for precise gene editing. However, the approaches for precise control over the genome editing and regulatory events are still desirable. Here, we reported a spatiotemporal and efficient CRISPR/Cas9 and Cpf1-mediated editing with photo-sensitive circular gRNAs. This approach relies on only two or three pre-installed photolabile substituents followed by a simple covalent cyclization, which provides a robust synthesize approach in comparison to heavily modified gRNAs. In established cells stably expressing Cas9, the circular gRNA in coordination with light irradiation could direct a precise cleavage of GFP and VEGFA within a pre-defined cutting region. We have also achieved light-mediated MSTN gene editing in embryos, whereas a new bow-knot-type gRNA showed no background editing in the absence of light irradiation. Together, our work provides a significantly improved method to precisely manipulate where and when genes are edited. | ||
| 700 | 1 | |a Liu, Ji |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Jun-Jin |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Yu |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Wen-Da |e verfasserin |4 aut | |
| 700 | 1 | |a Cai, Wei-Qi |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, Li |e verfasserin |4 aut | |
| 700 | 1 | |a Tang, Xin-Jing |e verfasserin |4 aut | |
| 700 | 1 | |a Hou, Jian |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Ming |e verfasserin |4 aut | |
| 700 | 1 | |a Cheng, Liang |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t bioRxiv.org |g (2022) vom: 20. Mai |
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