Engineering of Zinc Finger Nucleases Through Structural Modeling Improves Genome Editing Efficiency in Cells
Abstract: Genome Editing is widely used in biomedical research and medicine. Zinc finger nucleases (ZFNs) are smaller in size than transcription activator‐like effector (TALE) nucleases (TALENs) and CRISPR‐Cas9. Therefore, ZFN‐encoding DNAs can be easily packaged into a viral vector with limited cargo space, such as adeno‐associated virus (AAV) vectors, for in vivo and clinical applications. ZFNs have great potential for translational research and clinical use. However, constructing functional ZFNs and improving their genome editing efficiency is extremely difficult. Here, the efficient construction of functional ZFNs and the improvement of their genome editing efficiency using AlphaFold, Coot, and Rosetta are described. Plasmids encoding ZFNs consisting of six fingers using publicly available zinc‐finger resources are assembled. Two functional ZFNs from the ten ZFNs tested are successfully obtained. Furthermore, the engineering of ZFNs using AlphaFold, Coot, or Rosetta increases the efficiency of genome editing by 5%, demonstrating the effectiveness of engineering ZFNs based on structural modeling.
- Location
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Deutsche Nationalbibliothek Frankfurt am Main
- Extent
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Online-Ressource
- Language
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Englisch
- Bibliographic citation
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Engineering of Zinc Finger Nucleases Through Structural Modeling Improves Genome Editing Efficiency in Cells ; day:10 ; month:04 ; year:2024 ; extent:9
Advanced science ; (10.04.2024) (gesamt 9)
- Creator
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Katayama, Shota
Watanabe, Masahiro
Kato, Yoshio
Nomura, Wataru
Yamamoto, Takashi
- DOI
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10.1002/advs.202310255
- URN
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urn:nbn:de:101:1-2024041114410525067981
- Rights
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Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Last update
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11.01.2024, 8:12 AM CET
Data provider
Deutsche Nationalbibliothek. If you have any questions about the object, please contact the data provider.
Associated
- Katayama, Shota
- Watanabe, Masahiro
- Kato, Yoshio
- Nomura, Wataru
- Yamamoto, Takashi
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