Engineering TadA ortholog-derived cytosine base editor without motif preference and adenosine activity limitation
The engineered TadA variants used in cytosine base editors (CBEs) present distinctive 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 employ 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 achieves robust exon 55 skipping and restoration of dystrophin expression. Our advancement in engineering TadA ortholog for cytosine editing enriches the base editing toolkits for gene-editing therapy and other potential applications.
Authors
- Shuna Cui (ORCID: https://orcid.org/0000-0003-2011-0314)
- Haoqiang Wang (ORCID: https://orcid.org/0000-0003-4944-1117)
- Yingsi Zhou (ORCID: https://orcid.org/0000-0002-6541-2116)
- Tanglong Yuan (ORCID: https://orcid.org/0000-0003-4670-2625)
- Jinhai Huang (ORCID: https://orcid.org/0000-0001-9952-3175)
- Chunlong Xu (ORCID: https://orcid.org/0000-0003-3533-3465)
- Erwei Zuo (ORCID: https://orcid.org/0000-0001-8259-0275)
- Zhenhai Zeng (ORCID: https://orcid.org/0000-0002-3247-2788)
- Guoling Li (ORCID: https://orcid.org/0000-0003-0422-6157)
- Yuyang Yu (ORCID: https://orcid.org/0009-0007-1584-4985)
- Guoli Zhao (ORCID: https://orcid.org/0000-0002-7807-5980)
- Xue Dong (ORCID: https://orcid.org/0000-0002-3355-7476)
- Yuan Yuan (ORCID: https://orcid.org/0000-0001-7877-3175)
- Jingxing Zhou (ORCID: https://orcid.org/0000-0002-5458-5783)
- Hainan Zhang (ORCID: https://orcid.org/0000-0002-1058-0462)
- Jiamin Luo (ORCID: https://orcid.org/0000-0003-2829-1914)
- Tong Li (ORCID: https://orcid.org/0009-0001-3934-9184)
- Yin Wang
Institutions
- XLAB (Slovenia) (SI)
- Chinese Academy of Medical Sciences & Peking Union Medical College (CN)
- Agricultural Genomics Institute at Shenzhen (CN)
- Huadong Hospital (CN)
- Eye & ENT Hospital of Fudan University (CN)
- Chinese Academy of Agricultural Sciences (CN)
- NewGen Therapeutics (United States) (US)
- Ministry of Agriculture and Rural Affairs (CN)
Publication Details
- Journal
- Nature Communications
- Published
- 2024-09-16
- DOI
- https://doi.org/10.1038/s41467-024-52485-1
- Citations
- 7
- Primary Topic
- CRISPR and Genetic Engineering
- Type
- article
- Field-Weighted Citation Impact
- 1.64
Funders
- National Natural Science Foundation of China
- Government of Jiangsu Province
- Science and Technology Commission of Shanghai Municipality
- Chinese Academy of Agricultural Sciences
- Jiangsu Provincial Key Research and Development Program