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

Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion.
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DOI:
10.1186/s13059-018-1443-z
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发表时间:
2018-05-29
期刊:
影响因子:
12.3
通讯作者:
Gao C
Gao C
中科院分区:
生物学1区
文献类型:
--
作者:
Li C;Zong Y;Wang Y;Jin S;Zhang D;Song Q;Zhang R;Gao C

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植物中的核苷酸碱基编辑器仅限于将胞嘧啶转化为胸腺嘧啶。在这里,我们描述了一种新的植物腺嘌呤碱基编辑器,它基于与切口酶CRISPR/Cas9融合的进化的tRNA腺苷脱氨酶,使A·T转化为G·C的频率在原生质体中高达7.5%,在再生的水稻和小麦植物中高达59.1%。还通过引入功能获得性点突变成功地修饰了内源基因,以直接产生耐除草剂的水稻植物。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,从而扩展了植物精确编辑的工具集。本文的在线版本(10.1186/s13059-018-1443-z)包含补充材料,可供授权用户使用。
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. The online version of this article (10.1186/s13059-018-1443-z) contains supplementary material, which is available to authorized users.
完美匹配的 20 核苷酸引导 RNA 序列可使用高保真 SpCas9 核酸酶进行稳健的基因组编辑。
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