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TitleEngineered IscB-ωRNA system with improved base editing efficiency for disease correction via single AAV delivery in mice.
Journal, issue, pagesCell Rep, Vol. 43, Issue 11, Page 114973, Year 2024
Publish dateNov 26, 2024
AuthorsRuochen Guo / Xiaozhi Sun / Feizuo Wang / Dingyi Han / Qiaoxia Yang / Hua Gao / Zhifang Li / Zhuang Shao / Jinqi Shi / Rongrong Yang / Xiaona Huo / Junda Yan / Guoling Li / Qingquan Xiao / Yuanhua Liu / Senfeng Zhang / Xinyu Liu / Yingsi Zhou / Leyun Wang / Chunyi Hu / Chunlong Xu /
PubMed AbstractIscBs, as hypercompact ancestry proteins of Cas9 nuclease, are suitable for in vivo gene editing via single adeno-associated virus (AAV) delivery. Due to the low activity of natural IscBs in ...IscBs, as hypercompact ancestry proteins of Cas9 nuclease, are suitable for in vivo gene editing via single adeno-associated virus (AAV) delivery. Due to the low activity of natural IscBs in eukaryotic cells, recent studies have been focusing on improving OgeuIscB's gene editing efficiency via protein engineering. However, in vivo gene editing efficacy of IscBs for disease correction remained to be demonstrated. Here, we showed effective gene knockout and base editing in mouse embryos. To further improve IscB activity, we performed systematic engineering of IscB-associated ωRNA and identified a variant, ωRNA-v2, with enhanced gene editing efficiency. Furthermore, our study demonstrated the efficacy of an engineered IscB-ωRNA system for robust gene knockout and base editing in vivo. Single AAV delivery of IscB-derived cytosine and adenine base editors achieved disease correction in a mouse model of tyrosinemia. Therefore, our results indicated the great potential of miniature IscBs for developing single-AAV-based gene editing therapeutics.
External linksCell Rep / PubMed:39541214
MethodsEM (single particle)
Resolution6.5 Å
Structure data

EMDB-62146: Engineered IscB-Version 2 wRNA-Target DNA ternary complex(enIscB-v2 wRNA-target DNA)
Method: EM (single particle) / Resolution: 6.5 Å

Source
  • Escherichia coli (E. coli)

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