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在植物中使用基因编辑进行本地基因沉默

Rundong Shen^1,2,3, Qi Yao^1,3, Xinhang Tan³

¹Shanghai Collaborative Innovation Center of Agri-Seeds, Joint Center for Single Cell Biology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China.

The New phytologist

|May 27, 2024

概括

一种新型的基因沉默方法使用上游启动密码子含有元素 (ATGE) 来有效降低植物的基因表达. 这种可编程方法为作物改良和生物研究提供了强大的工具.

科学领域:

植物分子生物学植物分子生物学
基因调节基因调节
农作物科学农作物科学

背景情况:

基因沉默对于开发所需的作物特征至关重要.
目前的RNA干扰 (RNAi) 方法面临着转基因不稳定性等局限性.
在5'UTR中,上游开始编码子 (uATG) 可以抑制蛋白质翻译.

研究的目的:

开发一种高效,强大的植物基因沉默方法.
为了研究在植物基因中引入上游启动含子元素 (ATGEs) 的有效性.
为了使可调节的基因敲除用于作物改良和生物研究.

主要方法:

设计并将量身定制的ATGE引入植物目标基因的5'UTR.
使用基准编辑引入uATG,将其有效性与ATGE插入进行比较.
优化的ATGE设计,包括双向多功能ATGE4,用于可调节的基因淘汰.
将ATGE策略应用于参与粉合成的Waxy基因.

主要成果:

直接将ATGE插入到主起始编码子 (pATG) 旁边,有效降低了目标蛋白质的表达.
试图引入uATG的基编辑尝试在制目标基因方面取得了有限的成功.
该ATGE战略实现了一致和实质性的目标蛋白下调.
使用ATGE4.4实现了19%至89%的可调节基因敲除.
将ATGE引入Waxy基因导致谷物中的粉糖含量减少.

结论:

开发了一种可编程和强大的方法,用于使用ATGEs在植物中的基因敲除.
该ATGE策略克服了传统RNAi的局限性,提供了更大的控制和稳定性.
这种方法具有探索基因功能和增强作物特征的巨大潜力.

关键词:

下调调节的下调调节方法基因组编辑基因组编辑分子繁殖的分子繁殖.米米饭米饭米饭米饭.一个序列插入插入.

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