文献类型： 外文期刊
作者： Shen, Rundong 1 ; Yao, Qi 1 ; Tan, Xinhang 4 ; Ren, Wendan 1 ; Zhong, Dating 1 ; Zhang, Xuening 1 ; Li, Xinbo 2 ; Dong, Chao 2 ; Cao, Xuesong 5 ; Tian, Yifu 1 ; Zhu, Jian-Kang 2 ; Lu, Yuming 1 ;
作者机构： 1.Shanghai Jiao Tong Univ, Shanghai Collaborat Innovat Ctr Agriseeds, Joint Ctr Single Cell Biol, Sch Agr & Biol, Shanghai 200240, Peoples R China
2.Chinese Acad Agr Sci CAAS, Inst Crop Sci, Natl Nanfan Res Inst, Sanya 572024, Peoples R China
3.Minist Agr & Rural Affairs, Key Lab Gene Editing Technol Hainan, Sanya 572024, Peoples R China
4.Chinese Acad Sci, Shanghai Ctr Plant Stress Biol, Ctr Excellence Mol Plant Sci, Shanghai 201602, Peoples R China
5.Southern Univ Sci & Technol, Inst Adv Biotechnol, Shenzhen 518055, Peoples R China
6.Southern Univ Sci & Technol, Sch Life Sci, Shenzhen 518055, Peoples R China
关键词： downregulation; genome editing; molecular breeding; rice; sequence insertion
期刊名称：NEW PHYTOLOGIST （ 影响因子：9.4； 五年影响因子：10.5 ）
ISSN： 0028-646X
年卷期： 2024 年
页码：
收录情况： SCI
摘要： Gene silencing is crucial in crop breeding for desired trait development. RNA interference (RNAi) has been used widely but is limited by ectopic expression of transgenes and genetic instability. Introducing an upstream start codon (uATG) into the 5 ' untranslated region (5 ' UTR) of a target gene may 'silence' the target gene by inhibiting protein translation from the primary start codon (pATG). Here, we report an efficient gene silencing method by introducing a tailor-designed uATG-containing element (ATGE) into the 5 ' UTR of genes in plants, occupying the original start site to act as a new pATG. Using base editing to introduce new uATGs failed to silence two of the tested three rice genes, indicating complex regulatory mechanisms. Precisely inserting an ATGE adjacent to pATG achieved significant target protein downregulation. Through extensive optimization, we demonstrated this strategy substantially and consistently downregulated target protein expression. By designing a bidirectional multifunctional ATGE4, we enabled tunable knockdown from 19% to 89% and observed expected phenotypes. Introducing ATGE into Waxy, which regulates starch synthesis, generated grains with lower amylose, revealing the value for crop breeding. Together, we have developed a programmable and robust method to knock down gene expression in plants, with potential for biological mechanism exploration and crop enhancement.
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