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利用 Cas9-腺嘌呤脱氨酶融合蛋白在水稻和小麦中进行扩展碱基编辑。

Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion.

机构信息

State Key Laboratory of Plant Cell and Chromosome Engineering, Center for Genome Editing, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China.

University of Chinese Academy of Sciences, Beijing, China.

出版信息

Genome Biol. 2018 May 29;19(1):59. doi: 10.1186/s13059-018-1443-z.

Abstract

Nucleotide base editors in plants have been limited to conversion of cytosine to thymine. Here, we describe a new plant adenine base editor based on an evolved tRNA adenosine deaminase fused to the nickase CRISPR/Cas9, enabling A•T to G•C conversion at frequencies up to 7.5% in protoplasts and 59.1% in regenerated rice and wheat plants. An endogenous gene is also successfully modified through introducing a gain-of-function point mutation to directly produce an herbicide-tolerant rice plant. With this new adenine base editing system, it is now possible to precisely edit all base pairs, thus expanding the toolset for precise editing in plants.

摘要

在植物中,核苷酸碱基编辑器一直仅限于将胞嘧啶转换为胸腺嘧啶。在这里,我们描述了一种新的基于进化的 tRNA 腺苷脱氨酶与 Nickase CRISPR/Cas9 融合的植物腺嘌呤碱基编辑器,能够在原生质体中达到高达 7.5%的 A•T 到 G•C 转换频率,在再生的水稻和小麦植物中达到 59.1%。通过引入功能获得性点突变,还成功修饰了一个内源性基因,直接产生了一种耐受除草剂的水稻植物。有了这个新的腺嘌呤碱基编辑系统,现在可以精确编辑所有碱基对,从而扩展了植物中精确编辑的工具集。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e6ad/5972399/5476118e4ab3/13059_2018_1443_Fig1_HTML.jpg

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