4.8 Article

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

Journal

NANO LETTERS
Volume 21, Issue 13, Pages 5859-5866

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acs.nanolett.1c01792

Keywords

gold nanoclusters; siRNA; delivery; gene silencing; plants

Funding

  1. Burroughs Wellcome Fund Career Award at the Scientific Interface (CASI)
  2. Stanley Fahn PDF Junior Faculty Grant [PF-JFA-1760]
  3. Beckman Foundation Young Investigator Award
  4. USDA AFRI award
  5. Gordon and Betty Moore Foundation
  6. USDA NIFA award
  7. USDA-BBT EAGER award
  8. NSF CAREER award
  9. Dreyfus Foundation
  10. Chan-Zuckerberg foundation
  11. FFAR New Innovator Award
  12. Office of Science, Office of Basic Energy Sciences, of the U.S. Department of Energy [DE-AC02-05CH11231]
  13. Keck Foundation [89208]
  14. Jinan University [88016105]
  15. Foundation for Food and Agriculture Research Fellowship
  16. Schlumberger Foundation Faculty
  17. Resnick Sustainability Institute
  18. UC Berkeley Molecular Imaging Center (Gordon and Betty Moore Foundation)
  19. QB3 Shared Stem Cell Facility
  20. Innovative Genomics Institute (IGI)
  21. BASF-CARA program

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The study synthesizes PEI-AuNCs for delivering siRNA into intact plants, achieving efficient gene knockdown without being hindered by the plant cell wall, providing a new approach for broad applications in plant biotechnology.
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.

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