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用于植物中高效小干扰 RNA 递送和基因沉默的碳点。

Carbon Dots for Efficient Small Interfering RNA Delivery and Gene Silencing in Plants.

机构信息

Bayer Crop Science, Woodland, California 95695

Bayer Crop Science, Woodland, California 95695.

出版信息

Plant Physiol. 2020 Oct;184(2):647-657. doi: 10.1104/pp.20.00733. Epub 2020 Aug 6.

DOI:10.1104/pp.20.00733
PMID:32764133
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7536711/
Abstract

The initiation of RNA interference (RNAi) by topically applied small interfering RNA has potential applications for plant functional genomics, crop improvement and crop protection, but the primary obstacle for the development of this technology is the efficient delivery of RNAi effectors into the cell. The plant cell wall is a particularly challenging barrier for the delivery of macromolecules because many of the transfection agents that are commonly used with animal cells produce nanocomplexes that are significantly larger than the size exclusion limit of the cell wall. Here, we illustrate the use of a class of very small nanoparticles, called carbon dots, for delivering small interfering RNA into the model plants and tomato (). Low-pressure spray application of these formulations with a spreading surfactant resulted in strong silencing of transgenes in both species. The delivery efficacy of carbon dot formulations was also demonstrated by the silencing of endogenous genes that encode two subunits of magnesium chelatase, an enzyme necessary for chlorophyll synthesis. The strong visible phenotypes observed with the carbon dot-facilitated delivery were validated by measuring significant reductions in the target gene transcript and/or protein levels. Methods for the delivery of RNAi effectors into plants, such as the carbon dot formulations described here, could become valuable tools for gene silencing in plants with practical applications in plant functional genomics and agriculture.

摘要

RNA 干扰(RNAi)的启动通过局部应用小干扰 RNA 具有应用于植物功能基因组学、作物改良和作物保护的潜力,但该技术发展的主要障碍是将 RNAi 效应物有效递送至细胞内。植物细胞壁是大分子递送至细胞内的一个特别具有挑战性的障碍,因为许多常用于动物细胞的转染试剂产生的纳米复合物的尺寸明显大于细胞壁的排除极限。在这里,我们展示了一类非常小的纳米颗粒,称为碳点,用于将小干扰 RNA 递送至模式植物 和番茄()。用具有铺展表面活性剂的低压喷雾应用这些制剂导致两种物种中转基因的强烈沉默。碳点制剂的递送功效也通过沉默编码镁螯合酶两个亚基的内源性基因得到证明,该酶是叶绿素合成所必需的。通过测量靶基因转录物和/或蛋白水平的显著降低,验证了用碳点促进的递送来观察到的强可见表型。用于将 RNAi 效应物递送至植物的方法,如这里描述的碳点制剂,可能成为植物基因沉默的有价值的工具,在植物功能基因组学和农业中有实际应用。

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本文引用的文献

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