Gold-Nanocluster-Mediated Delivery of siRNA to Intact Plant Cells for Efficient Gene Knockdown,Nano Letters

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Gold-Nanocluster-Mediated Delivery of siRNA to Intact Plant Cells for Efficient Gene Knockdown
Nano Letters ( IF 9.6 ) Pub Date : 2021-06-21 , DOI: 10.1021/acs.nanolett.1c01792
Huan Zhang _{1,

2} , Yuhong Cao ₃ , Dawei Xu ₂ , Natalie S Goh ₂ , Gozde S Demirer ₂ , Stefano Cestellos-Blanco ₃ , Yuan Chen ₄ , Markita P Landry _{2,

5,

6} , Peidong Yang _{3,

7,

8,

9}

Affiliation

Department of Chemistry, College of Chemistry and Materials Science, Jinan University, Guangzhou, Guangdong 510632, People's Republic of China.
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, California 94720, United States.
Department of Materials Science and Engineering, University of California, Berkeley, California 94720, United States.
Plant Gene Expression Center, United States Department of Agriculture-Agricultural Research Service, and Department of Plant and Microbial Biology, University of California Berkeley, Albany, California 94710, United States.
California Institute for Quantitative Biosciences, QB3, University of California, Berkeley, California 94720, United States.
Chan Zuckerberg BioHub, San Francisco, California 94158, United States.
Department of Chemistry, University of California, Berkeley, California 94720, United States.
Chemical Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States.
Kavli Energy NanoScience Institute, University of California, Berkeley, California 94720, United States.

RNA interference, which involves the delivery of small interfering RNA (siRNA), has been used to validate target genes, to understand and control cellular metabolic pathways, and to use as a “green” alternative to confer pest tolerance in crops. Conventional siRNA delivery methods such as viruses and Agrobacterium-mediated delivery exhibit plant species range limitations and uncontrolled DNA integration into the plant genome. Here, we synthesize polyethylenimine-functionalized gold nanoclusters (PEI-AuNCs) to mediate siRNA delivery into intact plants and show that these nanoclusters enable efficient gene knockdown. We further demonstrate that PEI-AuNCs protect siRNA from RNase degradation while the complex is small enough to bypass the plant cell wall. Consequently, AuNCs enable gene knockdown with efficiencies of up 76.5 ± 5.9% and 76.1 ± 9.5% for GFP and ROQ1, respectively, with no observable toxicity. Our data suggest that AuNCs can deliver siRNA into intact plant cells for broad applications in plant biotechnology.

中文翻译：

金纳米簇介导的 siRNA 递送至完整植物细胞以实现有效的基因敲除

RNA 干扰涉及小干扰 RNA (siRNA) 的传递，已被用于验证靶基因、了解和控制细胞代谢途径，并用作赋予作物害虫耐受性的“绿色”替代方案。传统的 siRNA 递送方法（例如病毒和农杆菌介导的递送）表现出植物物种范围的限制以及不受控制的 DNA 整合到植物基因组中。在这里，我们合成了聚乙烯亚胺功能化金纳米簇（PEI-AuNC）来介导 siRNA 递送到完整植物中，并表明这些纳米簇能够实现有效的基因敲除。我们进一步证明，PEI-AuNCs 可以保护 siRNA 免受 RNase 降解，同时该复合物足够小，可以绕过植物细胞壁。因此，AuNC 对 GFP 和 ROQ1 的基因敲除效率分别高达 76.5 ± 5.9% 和 76.1 ± 9.5%，且没有可观察到的毒性。我们的数据表明 AuNC 可以将 siRNA 传递到完整的植物细胞中，在植物生物技术中得到广泛应用。

更新日期：2021-07-14

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