A Conformational Restriction Strategy for the Control of CRISPR/Cas Gene Editing with Photoactivatable Guide RNAs**
Abstract The CRISPR/Cas system is one of the most powerful tools for gene editing. However, approaches for precise control of genome editing and regulatory events are still desirable. Here, we report the spatiotemporal and efficient control of CRISPR/Cas9‐ and Cas12a‐mediated editing with conformationally restricted guide RNAs (gRNAs). This approach relied on only two or three pre‐installed photo‐labile substituents followed by an intramolecular cyclization, representing a robust synthetic method in comparison to the heavily modified linear gRNAs that often require extensive screening and time‐consuming optimization. This tactic could direct the precise cleavage of the genes encoding green fluorescent protein (GFP) and the vascular endothelial growth factor A (VEGFA) protein within a predefined cutting region without notable editing leakage in live cells. We also achieved light‐mediated myostatin ( MSTN) gene editing in embryos, wherein a new bow‐knot‐type gRNA was constructed with excellent OFF/ON switch efficiency. Overall, our work provides a significant new strategy in CRISPR/Cas editing with modified circular gRNAs to precisely manipulate where and when genes are edited..
Medienart: |
E-Artikel |
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Erscheinungsjahr: |
2023 |
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Erschienen: |
2023 |
Enthalten in: |
Zur Gesamtaufnahme - volume:62 |
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Enthalten in: |
Angewandte Chemie International Edition - 62(2023), 5 |
Beteiligte Personen: |
Sun, Ying‐Jie [VerfasserIn] |
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Anmerkungen: |
© 2023 Wiley‐VCH GmbH |
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Umfang: |
10 |
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doi: |
10.1002/anie.202212413 |
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funding: |
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Förderinstitution / Projekttitel: |
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PPN (Katalog-ID): |
WLY015042448 |
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520 | |a Abstract The CRISPR/Cas system is one of the most powerful tools for gene editing. However, approaches for precise control of genome editing and regulatory events are still desirable. Here, we report the spatiotemporal and efficient control of CRISPR/Cas9‐ and Cas12a‐mediated editing with conformationally restricted guide RNAs (gRNAs). This approach relied on only two or three pre‐installed photo‐labile substituents followed by an intramolecular cyclization, representing a robust synthetic method in comparison to the heavily modified linear gRNAs that often require extensive screening and time‐consuming optimization. This tactic could direct the precise cleavage of the genes encoding green fluorescent protein (GFP) and the vascular endothelial growth factor A (VEGFA) protein within a predefined cutting region without notable editing leakage in live cells. We also achieved light‐mediated myostatin ( MSTN) gene editing in embryos, wherein a new bow‐knot‐type gRNA was constructed with excellent OFF/ON switch efficiency. Overall, our work provides a significant new strategy in CRISPR/Cas editing with modified circular gRNAs to precisely manipulate where and when genes are edited. | ||
700 | 1 | |a Chen, Wen‐Da |4 aut | |
700 | 1 | |a Liu, Ji |4 aut | |
700 | 1 | |a Li, Jun‐Jin |4 aut | |
700 | 1 | |a Zhang, Yu |4 aut | |
700 | 1 | |a Cai, Wei‐Qi |4 aut | |
700 | 1 | |a Liu, Li |4 aut | |
700 | 1 | |a Tang, Xin‐Jing |4 aut | |
700 | 1 | |a Hou, Jian |4 aut | |
700 | 1 | |a Wang, Ming |4 aut | |
700 | 1 | |a Cheng, Liang |4 aut | |
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