DNA nanostructures coordinate gene silencing in mature plants
DNA nanostructures coordinate gene silencing in mature plants
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DNA纳米结构协调成熟植物中的基因沉默
DOI：
10.1073/pnas.1818290116
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发表时间：
2019-04-09
影响因子：
11.1
通讯作者：
Landry, Markita P.
中科院分区：
文献类型：
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作者：
Zhang, Huan;Demirer, Gozde S.;Landry, Markita P.
向植物传递生物分子依赖于农杆菌感染或生物颗粒传递，前者仅适用于DNA传递。将RNA等功能性生物分子传递到植物细胞的困难是由于植物细胞壁，它在哺乳动物细胞中是不存在的，并且构成了植物生物分子传递的主要物理屏障。DNA纳米结构介导的生物分子递送是一种有效的跨哺乳动物细胞脂双层递送的策略；然而，在植物中，没有外部机械辅助的纳米颗粒介导的生物分子递送仍未被探索。在此，我们对不同的DNA纳米结构进行了系统的评估，以了解它们内化到成熟植物细胞中的能力，传递siRNAs，并有效地沉默烟草叶片中一个结构性表达的基因。我们发现，纳米结构内化到植物细胞中以及相应的基因沉默效率取决于DNA纳米结构的大小、形状、致密性、硬度和siRNA连接位点在纳米结构上的位置。我们进一步证实，DNA纳米结构的内化效率与它们各自的基因沉默效率有关，但内源基因沉默途径依赖于siRNA连接位点。我们的工作建立了将生物分子输送到具有DNA纳米结构的植物中的可行性，详细介绍了植物细胞内化的重要设计参数，并评估了DNA纳米结构几何形状对基因沉默机制的影响。
Delivery of biomolecules to plants relies on Agrobacterium infection or biolistic particle delivery, the former of which is amenable only to DNA delivery. The difficulty in delivering functional biomolecules such as RNA to plant cells is due to the plant cell wall, which is absent in mammalian cells and poses the dominant physical barrier to biomolecule delivery in plants. DNA nanostructure-mediated biomolecule delivery is an effective strategy to deliver cargoes across the lipid bilayer of mammalian cells; however, nanoparticle-mediated delivery without external mechanical aid remains unexplored for biomolecule delivery across the cell wall in plants. Herein, we report a systematic assessment of different DNA nanostructures for their ability to internalize into cells of mature plants, deliver siRNAs, and effectively silence a constitutively expressed gene in Nicotiana benthamiana leaves. We show that nanostructure internalization into plant cells and corresponding gene silencing efficiency depends on the DNA nanostructure size, shape, compactness, stiffness, and location of the siRNA attachment locus on the nanostructure. We further confirm that the internalization efficiency of DNA nanostructures correlates with their respective gene silencing efficiencies but that the endogenous gene silencing pathway depends on the siRNA attachment locus. Our work establishes the feasibility of biomolecule delivery to plants with DNA nanostructures and both details the design parameters of importance for plant cell internalization and also assesses the impact of DNA nanostructure geometry for gene silencing mechanisms.