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
Deutsche Nationalbibliothek Frankfurt am Main
Extent
Online-Ressource
Language
Englisch

Bibliographic citation
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
Katayama, Shota
Watanabe, Masahiro
Kato, Yoshio
Nomura, Wataru
Yamamoto, Takashi

DOI
10.1002/advs.202310255
URN
urn:nbn:de:101:1-2024041114410525067981
Rights
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
Last update
11.01.2024, 8:12 AM CET

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Associated

  • Katayama, Shota
  • Watanabe, Masahiro
  • Kato, Yoshio
  • Nomura, Wataru
  • Yamamoto, Takashi

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