设计微型 IscB 标记酶，实现稳健的碱基编辑和广泛的靶向范围

IF 12.9 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Nature chemical biology Pub Date : 2024-07-08 DOI:10.1038/s41589-024-01670-w

Linxiao Han, Yueer Hu, Qiqin Mo, Hao Yang, Feng Gu, Fang Bai, Yadong Sun, Hanhui Ma

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引用次数: 0

摘要

IscB与Cas9具有相似的结构域，但IscB体积小，更适合通过腺相关病毒传递。为了改善OgeuIscB（来自人类肠道元基因组的IscB）在哺乳动物细胞中的低编辑效率，我们通过工程化OgeuIscB缺口酶及其同源ωRNA（称为IminiBEs），开发了高效微型碱基编辑器。我们证明了 IminiCBE（平均 67%）或 IminiABE（平均 52%）的强大编辑效率。将非特异性 DNA 结合蛋白 Sso7d 融合到 IminiBEs 中可将低效位点的编辑效率提高约两到三倍，我们称之为 SIminiBEs。此外，IminiCBE 和 SIminiCBE 还能识别 NNRR、NNRY 和 NNYR 目标相邻基序，从而拓宽了野生型 IscB 缺口酶的 NWRRNA 目标相邻基序位点。总之，IminiBEs 和 SIminiBEs 是高效的微型碱基编辑器，可用于特定位点的基因组突变。

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Engineering miniature IscB nickase for robust base editing with broad targeting range

IscB has a similar domain organization to Cas9, but the small size of IscB is better suited for delivery by adeno-associated virus. To improve the low editing efficiency of OgeuIscB (IscB from human gut metagenome) in mammalian cells, we developed high-efficiency miniature base editors by engineering OgeuIscB nickase and its cognate ωRNA, termed IminiBEs. We demonstrated the robust editing efficiency of IminiCBE (67% on average) or IminiABE (52% on average). Fusing non-specific DNA-binding protein Sso7d to IminiBEs increased the editing efficiency of low-efficiency sites by around two- to threefold, and we termed it SIminiBEs. In addition, IminiCBE and SIminiCBE recognize NNRR, NNRY and NNYR target-adjacent motifs, which broaden the canonical NWRRNA target-adjacent motif sites for the wild-type IscB nickase. Overall, IminiBEs and SIminiBEs are efficient miniature base editors for site-specific genomic mutations.

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来源期刊

Nature chemical biology 生物-生化与分子生物学

CiteScore

23.90

自引率

1.40%

发文量

238

审稿时长

12 months

期刊介绍： Nature Chemical Biology stands as an esteemed international monthly journal, offering a prominent platform for the chemical biology community to showcase top-tier original research and commentary. Operating at the crossroads of chemistry, biology, and related disciplines, chemical biology utilizes scientific ideas and approaches to comprehend and manipulate biological systems with molecular precision. The journal embraces contributions from the growing community of chemical biologists, encompassing insights from chemists applying principles and tools to biological inquiries and biologists striving to comprehend and control molecular-level biological processes. We prioritize studies unveiling significant conceptual or practical advancements in areas where chemistry and biology intersect, emphasizing basic research, especially those reporting novel chemical or biological tools and offering profound molecular-level insights into underlying biological mechanisms. Nature Chemical Biology also welcomes manuscripts describing applied molecular studies at the chemistry-biology interface due to the broad utility of chemical biology approaches in manipulating or engineering biological systems. Irrespective of scientific focus, we actively seek submissions that creatively blend chemistry and biology, particularly those providing substantial conceptual or methodological breakthroughs with the potential to open innovative research avenues. The journal maintains a robust and impartial review process, emphasizing thorough chemical and biological characterization.

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