C-terminal frameshift mutations generate viable knockout mutants with developmental defects for three essential protein kinases

IF 4.6 4区 农林科学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Yun Zhang, Miao-Miao Cui, Run-Nan Ke, Yue-Dan Chen, Kabin Xie
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引用次数: 0

Abstract

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 MITOGEN-ACTIVATED PROTEIN KINASES 1 (OsMPK1) 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.

C 端框移突变可产生具有发育缺陷的可行基因敲除突变体,这三种蛋白激酶至关重要
功能缺失突变体是基因功能研究的基本资源。然而,要产生可行且可遗传的重要基因敲除突变体却很困难。在这里,我们发现靶向编辑胚胎致死基因 MITOGEN-ACTIVATED PROTEIN KINASES 1(OsMPK1)的 C 端序列可产生弱突变体。这种C端编辑的osmpk1突变体表现出严重的发育缺陷和抗病性改变,但却能产生数十粒继承了突变基因的可存活种子。利用相同的 C 端编辑方法,我们还获得了一种壁相关蛋白激酶(Os07g0493200)和一种富亮氨酸重复受体样蛋白激酶(Os01g0239700)的可存活突变体,而这些基因的无效突变是致死的。这些数据表明,蛋白激酶的活性可以通过引入C端附近的移帧突变来降低,这可以为基因功能研究提供宝贵的资源,并为信号通路工程调控蛋白激酶的活性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
7.70
自引率
2.80%
发文量
0
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