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Efficient Transient Gene Knock-down in Tobacco Plants Using Carbon Nanocarriers

Demirer, Gozde S. and Landry, Markita P. (2021) Efficient Transient Gene Knock-down in Tobacco Plants Using Carbon Nanocarriers. Bio-protocol, 11 (1). Art. No. e3897. ISSN 2331-8325. PMCID PMC7953349. doi:10.21769/bioprotoc.3897. https://resolver.caltech.edu/CaltechAUTHORS:20220302-323885000

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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.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://dx.doi.org/10.21769/BioProtoc.3897DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7953349/PubMed CentralArticle
ORCID:
AuthorORCID
Demirer, Gozde S.0000-0002-3007-1489
Landry, Markita P.0000-0002-5832-8522
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.
Group:Resnick Sustainability Institute
Funders:
Funding AgencyGrant Number
Schlumberger FoundationUNSPECIFIED
Resnick Sustainability InstituteUNSPECIFIED
Burroughs Wellcome FundUNSPECIFIED
Parkinson's FoundationPFJFA-1760
Arnold and Mabel Beckman FoundationUNSPECIFIED
Department of AgricultureUNSPECIFIED
Gordon and Betty Moore FoundationUNSPECIFIED
Foundation for Food ResearchUNSPECIFIED
Chan-Zuckerberg BiohubUNSPECIFIED
Subject Keywords:Plant gene silencing, RNA interference, siRNA delivery, Gene knock-down, Nicotiana benthamiana, Carbon nanotubes, Single-walled carbon nanotubes (SWNT), Nanomaterial
Issue or Number:1
PubMed Central ID:PMC7953349
DOI:10.21769/bioprotoc.3897
Record Number:CaltechAUTHORS:20220302-323885000
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20220302-323885000
Official Citation:Demirer, G. S., & Landry, M. P. (2021). Efficient Transient Gene Knock-down in Tobacco Plants Using Carbon Nanocarriers. Bio-protocol, 11(1), e3897. https://doi.org/10.21769/BioProtoc.3897
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:113701
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:03 Mar 2022 17:23
Last Modified:03 Mar 2022 17:23

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