Carbon Dots for Efficient Small Interfering RNA Delivery and Gene Silencing in Plants
Carbon Dots for Efficient Small Interfering RNA Delivery and Gene Silencing in Plants
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DOI：
10.1104/pp.20.00733
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发表时间：
2020-10-01
期刊：
影响因子：
7.4
通讯作者：
Zheng, Wei
中科院分区：
文献类型：
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作者：
Schwartz, Steven H.;Hendrix, Bill;Zheng, Wei
由局部应用的小干扰RNA引发的RNA干扰(RNAi)在植物功能基因组学、作物改良和作物保护方面具有潜在的应用前景，但这项技术发展的主要障碍是将RNAi效应子高效地输送到细胞内。植物细胞壁是大分子传递的一个特别具有挑战性的障碍，因为许多动物细胞常用的转染剂产生的纳米复合体明显大于细胞壁的大小排除限制。在这里，我们说明了一类非常小的纳米颗粒的使用，称为碳点，用于将小干扰RNA传递到模式植物烟草和番茄(Solanum Lycpersicum)中。将这些配方与扩散表面活性剂一起进行低压喷雾，可使两个物种中的GFP转基因发生强烈的沉默。碳点制剂的传递效率也通过沉默编码镁螯合酶两个亚单位的内源基因来证明，镁螯合酶是合成叶绿素所必需的一种酶。通过测量目标基因转录本和/或蛋白质水平的显著降低，验证了碳点促进交付中观察到的强烈可见表型。将RNAi效应器导入植物的方法，如本文所述的碳点制剂，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具有实际应用。
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 Nicotiana benthamiana and tomato (Solanum lycopersicum). Low-pressure spray application of these formulations with a spreading surfactant resulted in strong silencing of GFP 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.