High performance TadA-8e derived cytosine and dual base editors with undetectable off-target effects in plants

Tingting Fan, Yanhao Cheng, Yuechao Wu, Shishi Liu, Xu Tang, Yao He, Shanyue Liao, Xuelian Zheng, Tao Zhang (), Yiping Qi () and Yong Zhang ()
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Tingting Fan: University of Electronic Science and Technology of China
Yanhao Cheng: University of Maryland
Yuechao Wu: Yangzhou University
Shishi Liu: University of Electronic Science and Technology of China
Xu Tang: Southwest University
Yao He: University of Electronic Science and Technology of China
Shanyue Liao: University of Electronic Science and Technology of China
Xuelian Zheng: Southwest University
Tao Zhang: Yangzhou University
Yiping Qi: University of Maryland
Yong Zhang: Southwest University

Nature Communications, 2024, vol. 15, issue 1, 1-15

Abstract: Abstract Cytosine base editors (CBEs) and adenine base editors (ABEs) enable precise C-to-T and A-to-G edits. Recently, ABE8e, derived from TadA-8e, enhances A-to-G edits in mammalian cells and plants. Interestingly, TadA-8e can also be evolved to confer C-to-T editing. This study compares engineered CBEs derived from TadA-8e in rice and tomato cells, identifying TadCBEa, TadCBEd, and TadCBEd_V106W as efficient CBEs with high purity and a narrow editing window. A dual base editor, TadDE, promotes simultaneous C-to-T and A-to-G editing. Multiplexed base editing with TadCBEa and TadDE is demonstrated in transgenic rice, with no off-target effects detected by whole genome and transcriptome sequencing, indicating high specificity. Finally, two crop engineering applications using TadDE are shown: introducing herbicide resistance alleles in OsALS and creating synonymous mutations in OsSPL14 to resist OsMIR156-mediated degradation. Together, this study presents TadA-8e derived CBEs and a dual base editor as valuable additions to the plant editing toolbox.

Date: 2024
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DOI: 10.1038/s41467-024-49473-w

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