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Cited by 9CRISPR and Genetic EngineeringRNA regulation and diseaseVirus-based gene therapy research

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Engineering TadA ortholog-derived cytosine base editor without motif preference and adenosine activity limitation

Open access
Li, Guoling, Dong, Xue, Luo, Jiamin, Yuan, Tanglong, Li, Tong, Zhao, Guoli, Zhang, Hainan, Zhou, Jingxing, Zeng, Zhenhai, Cui, Shuna, Wang, Haoqiang, Wang, Yin, Yu, Yuyang, Yuan, Yuan, Zuo, Erwei, Xu, Chunlong, Huang, Jinhai, Zhou, Yingsi·2024·Nature Communications
DOI 10.1038/s41467-024-52485-1PMC PMC11405849

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Abstract

Abstract The engineered TadA variants used in cytosine base editors (CBEs) present unique advantages, including a smaller size and fewer off-target effects compared to cytosine base editors that rely on natural deaminases. However, the current TadA variants demonstrate a preference for base editing in DNA with specific motif sequences and possess dual deaminase activity, acting on both cytosine and adenosine in adjacent positions, limiting their application scope. To address these issues, we employed TadA orthologs screening and multi sequence alignment (MSA)-guided protein engineering techniques to create a highly effective cytosine base editor (aTdCBE) without motif and adenosine deaminase activity limitations. Notably, the delivery of aTdCBE to a humanized mouse model of Duchenne muscular dystrophy (DMD) mice achieved robust exon 55 skipping and restoration of dystrophin expression. Our advancement in engineering TadA ortholog for cytosine editing enrich the base editing toolkits for gene-editing therapy and other potential applications.

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Metadata

Authors
Li, G., Dong, X., Luo, J., Yuan, T., Li, T., Zhao, G., Zhang, H., Zhou, J., Zeng, Z., Cui, S., Wang, H., Wang, Y., Yu, Y., Yuan, Y., Zuo, E., Xu, C., Huang, J., & Zhou, Y.
Affiliations
  • Agricultural Genomics Institute at Shenzhen (CN)
  • Chinese Academy of Medical Sciences & Peking Union Medical College (CN)
Corresponding author
  • Zuo, Erwei
  • Xu, Chunlong
  • Huang, Jinhai
  • Zhou, Yingsi
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2024-09-16
Volume
15
Issue
1
Pages
8090-8090
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-024-52485-1
PMC
PMC11405849
OpenAlex
W4402581033
Cited by
9
References
8
Citations by year
2026: 2 · 2025: 7
FWCI
1.23
Type
journal article
Language
en
Update notices
correction (2024-10-18)
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by-nc-nd)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • National Natural Science Foundation of China (82301215, 82271048)
  • Science and Technology Commission of Shanghai Municipality (23XD1420500)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
Keywords
TADA, cytosine base editors, adenosine deaminase activity, protein engineering, exon skipping, Duchenne muscular dystrophy, dystrophin expression, gene editing therapy, off-target effects
MeSH terms
Gene Editing, Adenosine, Adenosine Deaminase, Animals, Cytosine, Disease Models, Animal, Exons, Humans, Protein Engineering, Dystrophin, Muscular Dystrophy, Duchenne, Escherichia coli Proteins, Mice, HEK293 Cells, CRISPR-Cas Systems

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