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- Title
Development of deaminase-free T-to-S base editor and C-to-G base editor by engineered human uracil DNA glycosylase.
- Authors
Tong, Huawei; Wang, Haoqiang; Wang, Xuchen; Liu, Nana; Li, Guoling; Wu, Danni; Li, Yun; Jin, Ming; Li, Hengbin; Wei, Yinghui; Li, Tong; Yuan, Yuan; Shi, Linyu; Yao, Xuan; Zhou, Yingsi; Yang, Hui
- Abstract
DNA base editors enable direct editing of adenine (A), cytosine (C), or guanine (G), but there is no base editor for direct thymine (T) editing currently. Here we develop two deaminase-free glycosylase-based base editors for direct T editing (gTBE) and C editing (gCBE) by fusing Cas9 nickase (nCas9) with engineered human uracil DNA glycosylase (UNG) variants. By several rounds of structure-informed rational mutagenesis on UNG in cultured human cells, we obtain gTBE and gCBE with high activity of T-to-S (i.e., T-to-C or T-to-G) and C-to-G conversions, respectively. Furthermore, we conduct parallel comparison of gTBE/gCBE with those recently developed using other protein engineering strategies, and find gTBE/gCBE show the outperformance. Thus, we provide several base editors, gTBEs and gCBEs, with corresponding engineered UNG variants, broadening the targeting scope of base editors. Efficient base editors for direct thymine (T) editing are highly desirable. Here, authors develop two deaminase-free glycosylase-based base editors for direct T editing (gTBE) and C editing (gCBE) by rounds of structure-informed mutagenesis on human DNA glycosylase UNG and further engineering.
- Subjects
HUMAN DNA; ADENINE; PROTEIN engineering; THYMINE; GUANINE; CYTOSINE
- Publication
Nature Communications, 2024, Vol 15, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-024-49343-5