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-te...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 wallassociated 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.展开更多
基金supported by the National Natural Science Foundation of China(32293243)Fundamental Research Funds for the Central Universities(2021ZKPY002,2662023PY006)supported by Hainan Yazhou Bay Seed Laboratory and the China National Seed Group(project B23YQ1516).
文摘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 wallassociated 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.
