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. 2023 Jul;20(7):1029-1036.
doi: 10.1038/s41592-023-01898-9. Epub 2023 May 25.

Development of miniature base editors using engineered IscB nickase

Dingyi Han #  1   2 , Qingquan Xiao #  1   3   4 , Yifan Wang #  1   2 , Hainan Zhang #  3   4 , Xue Dong  3 , Guoling Li  3 , Xiangfeng Kong  3   4 , Shihao Wang  3 , Jinhui Song  3   4 , Weihong Zhang  3   4 , Jingxing Zhou  3   4 , Lanting Bi  3   4 , Yuan Yuan  3 , Linyu Shi  3 , Na Zhong  3   4 , Hui Yang  5   6   7 , Yingsi Zhou  8   9
Affiliations

Development of miniature base editors using engineered IscB nickase

Dingyi Han et al. Nat Methods. 2023 Jul.

Abstract

As a miniature RNA-guided endonuclease, IscB is presumed to be the ancestor of Cas9 and to share similar functions. IscB is less than half the size of Cas9 and thus more suitable for in vivo delivery. However, the poor editing efficiency of IscB in eukaryotic cells limits its in vivo applications. Here we describe the engineering of OgeuIscB and its corresponding ωRNA to develop an IscB system that is highly efficient in mammalian systems, named enIscB. By fusing enIscB with T5 exonuclease (T5E), we found enIscB-T5E exhibited comparable targeting efficiency to SpG Cas9 while showing reduced chromosome translocation effects in human cells. Furthermore, by fusing cytosine or adenosine deaminase with enIscB nickase, we generated miniature IscB-derived base editors (miBEs), exhibiting robust editing efficiency (up to 92%) to induce DNA base conversions. Overall, our work establishes enIscB-T5E and miBEs as versatile tools for genome editing.

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