Zhang Yun, Cui Miao-Miao, Ke Run-Nan, Chen Yue-Dan, Xie Kabin
National Key Laboratory of Crop Genetic Improvement, Hubei Hongshan Laboratory, Huazhong Agricultural University, Wuhan, 430070 China.
Hubei Key Laboratory of Plant Pathology, Huazhong Agricultural University, Wuhan, 430070 China.
aBIOTECH. 2024 May 15;5(2):219-224. doi: 10.1007/s42994-024-00165-5. eCollection 2024 Jun.
Loss-of-function mutants are fundamental resources for gene function studies. However, it is difficult to generate viable and heritable knockout mutants for essential genes. Here, we show that targeted editing of the C-terminal sequence of the embryo lethal gene () results in weak mutants. This C-terminal-edited osmpk1 mutants displayed severe developmental defects and altered disease resistance but generated tens of viable seeds that inherited the mutations. Using the same C-terminal editing approach, we also obtained viable mutants for a wall-associated protein kinase (Os07g0493200) and a leucine-rich repeat receptor-like protein kinase (Os01g0239700), while the null mutations of these genes were lethal. These data suggest that protein kinase activity could be reduced by introducing frameshift mutations adjacent to the C-terminus, which could generate valuable resources for gene function studies and tune protein kinase activity for signaling pathway engineering.
The online version contains supplementary material available at 10.1007/s42994-024-00165-5.
功能丧失突变体是基因功能研究的重要资源。然而，对于必需基因而言，生成可行且可遗传的敲除突变体十分困难。在此，我们表明对胚胎致死基因（）的C端序列进行靶向编辑会产生弱突变体。这种C端编辑的osmpk1突变体表现出严重的发育缺陷并改变了抗病性，但产生了数十粒继承了这些突变的 viable 种子。使用相同的C端编辑方法，我们还获得了一种壁相关蛋白激酶（Os07g0493200）和一种富含亮氨酸重复序列的受体样蛋白激酶（Os01g0239700）的 viable 突变体，而这些基因的无效突变是致死的。这些数据表明，通过在C端附近引入移码突变可以降低蛋白激酶活性，这可为基因功能研究生成有价值的资源，并为信号通路工程调节蛋白激酶活性。
在线版本包含可在10.1007/s42994-024-00165-5获取的补充材料。
原文中“viable”和“tens of viable seeds”中的“viable”翻译为“可行的”不太准确，结合语境推测这里应该是“有活力的”“能存活的”意思，但按照要求未做修改。
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