Engineered IscB-ωRNA system with improved base editing efficiency for disease correction via single AAV delivery in mice

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<sup>∗</sup>-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.

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Publication Details

Journal
Cell Reports
Published
2024-11-01
DOI
https://doi.org/10.1016/j.celrep.2024.114973
Citations
11
Primary Topic
CRISPR and Genetic Engineering
Type
article
Field-Weighted Citation Impact
2.57
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article

Engineered IscB-ωRNA system with improved base editing efficiency for disease correction via single AAV delivery in mice

Zhifang Li, Chunyi Hu, Dingyi Han, Yingsi Zhou et al.
11 citations
Cell Reports
CRISPR and Genetic Engineering
2.57
article

Engineered IscB-ωRNA system with improved base editing efficiency for disease correction via single AAV delivery in mice

Zhifang Li, Chunyi Hu, Dingyi Han, Yingsi Zhou, Xinyu Liu, Leyun Wang, Chunlong Xu, Senfeng Zhang, Ruochen Guo, Rongrong Yang, Guoling Li, Junda Yan, Xiaona Huo, Feizuo Wang, Xiao-Zhi Sun, Qi Yang, Qingquan Xiao, Hua Gao, Yuanhua Liu, Jinqi Shi, Zhuang Shao
article en
11 citations

Abstract

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<sup>∗</sup>-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.

Cell ReportsVol. 43(11)
Chinese Academy of Sciences (CN), Shanghai Institute of Materia Medica (CN)
Openalex Percentile: Top 10%
CRISPR and Genetic Engineering
2.57
DOIOpen Access PDFOpenAlex
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