Efficient Transient Gene Knock-down in Tobacco Plants Using Carbon Nanocarriers
- Creators
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Demirer, Gozde S.
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Landry, Markita P.
Abstract
Gene knock-down in plants is a useful approach to study genotype-phenotype relationships, render disease resistance to crops, and enable efficient biosynthesis of molecules in plants. Small interfering RNA (siRNA)-mediated gene silencing is one of the most common ways to achieve gene knock-down in plants. Traditionally, siRNA is delivered into intact plant cells by coding the siRNA sequences into DNA vectors, which are then delivered through viral and/or bacterial methods. In this protocol, we provide an alternative direct delivery method of siRNA molecules into intact plant cells for efficient transient gene knock-down in model tobacco plant, Nicotiana benthamiana, leaves. Our approach uses one dimensional carbon-based nanomaterials, single-walled carbon nanotubes (SWNTs), to deliver siRNA, and does not rely on viral/bacterial delivery. The distinct advantages of our method are i) there is no need for DNA coding of siRNA sequences, ii) this abiotic method could work in a broader range of plant species than biotic methods, and iii) there are fewer regulatory complications when using abiotic delivery methods, whereby gene silencing is transient without permanent modification of the plant genome.
Additional Information
© 2021 The Authors; exclusive licensee Bio-protocol LLC. This protocol is derived from Demirer et al. (2020). G.S.D. is supported by the Schlumberger Foundation Faculty for the Future Program and Caltech Resnick Sustainability Institute. We acknowledge support of a Burroughs Wellcome Fund Career Award at the Scientific Interface (CASI), a Stanley Fahn PDF Junior Faculty Grant with Award # PFJFA-1760, a Beckman Foundation Young Investigator Award, a USDA AFRI award, a USDA NIFA award, the Moore Foundation, and an FFAR New Innovator Award (M.P.L). M.P.L. is a Chan-Zuckerberg Biohub investigator. Authors declare no competing interest.Attached Files
Published - BioProtoc-11-01-3897.pdf
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Additional details
- PMCID
- PMC7953349
- Eprint ID
- 113701
- Resolver ID
- CaltechAUTHORS:20220302-323885000
- Schlumberger Foundation
- Resnick Sustainability Institute
- Burroughs Wellcome Fund
- Parkinson's Foundation
- PFJFA-1760
- Arnold and Mabel Beckman Foundation
- Department of Agriculture
- Gordon and Betty Moore Foundation
- Foundation for Food Research
- Chan-Zuckerberg Biohub
- Created
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2022-03-03Created from EPrint's datestamp field
- Updated
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2022-03-03Created from EPrint's last_modified field
- Caltech groups
- Resnick Sustainability Institute