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Accelerated discovery and miniaturization of novel single-stranded cytidine deaminases

Nucleic Acids Research. 2024-09; 
Jiacheng Deng,† , Xueyuan Li,† , Hao Yu,† , Lin Yang1, Ziru Wang1, Wenfeng Yi, Ying Liu, Wenyu Xiao, Hongyong Xiang, Zicong Xie, Dongmei Lv, Hongsheng Ouyang, Daxin Pang, * and Hongming Yuan
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Gene Synthesis Following the construction strategy, 12 candidates were identified and synthesized with eukaryotic expression optimization by Genscript (China). Get A Quote

摘要

Cytidine base editors (CBEs) hold significant potential in genetic disease treatment and in breeding superior traits into animals. However, their large protein sizes limit their delivery by adeno-associated virus (AAV), given its packing capacity of <4.7 kb. To overcome this, we employed a web-based fast generic discovery (WFG) strategy, identifying several small ssDNA deaminases (Sdds) and constructing multiple Sdd-CBE 1.0 versions. SflSdd-CBE 1.0 demonstrated high C-to-T editing efficiency, comparable to AncBE4max, while SviSdd-CBE 1.0 exhibited moderate Cto-T editing efficiency with a narrow editing window (C3 to C5). Utilizing AlphaFold2, we devised a one-step miniaturization strategy, reducing the siz... More

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