scholarly journals Carbon dots for efficient siRNA delivery and gene silencing in plants

2019 ◽  
Author(s):  
Steven. H. Schwartz ◽  
Bill Hendrix ◽  
Paul Hoffer ◽  
Rick A. Sanders ◽  
Wei Zheng
Keyword(s):  
Cell Wall ◽  
Gene Silencing ◽  
Functional Genomics ◽  
Plant Cell ◽  
Carbon Dots ◽  
Plant Cell Wall ◽  
Crop Improvement ◽  
Size Exclusion ◽  
Gene Transcript ◽  
Carbon Dot

SUMMARYThe Initiation of RNA interference (RNAi) by topically applied double stranded RNA (dsRNA) has potential applications for plant functional genomics, crop improvement and crop protection. The primary obstacle for the development of this technology is efficient delivery of RNAi effectors. The plant cell wall is a particularly challenging barrier to the delivery of macromolecules. Many of the transfection agents that are commonly used with animal cells produce nanocomplexes that are significantly larger than the size exclusion limit of the plant cell wall. Utilizing a class of very small nanoparticles called carbon dots, a method of delivering siRNA into the model plant Nicotiana benthamiana and tomato is described. Low-pressure spray application of these formulations with a spreading surfactant resulted in strong silencing of GFP transgenes in both species. The delivery efficacy of carbon dot formulations was also demonstrated by silencing endogenous genes that encode two sub-units of magnesium chelatase, an enzyme necessary for chlorophyll synthesis. The strong visible phenotypes observed with the carbon dot facilitated delivery were validated by measuring significant reductions in the target gene transcript and/or protein levels. Methods for the delivery of RNAi effectors into plants, such as the carbon dot formulations described here, could become valuable tools for gene silencing in plants with practical applications in plant functional genomics and agriculture.

scholarly journals Virus-Induced Gene Silencing Offers a Functional Genomics Platform for Studying Plant Cell Wall Formation

2010 ◽  
Vol 3 (5) ◽  
pp. 818-833 ◽  
Author(s):  
Xiaohong Zhu ◽  
Sivakumar Pattathil ◽  
Koushik Mazumder ◽  
Amanda Brehm ◽  
Michael G. Hahn ◽  
...  
Keyword(s):  
Cell Wall ◽  
Gene Silencing ◽  
Functional Genomics ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Cell Wall Formation ◽  
Virus Induced Gene Silencing ◽  
Wall Formation

scholarly journals DNA nanostructures coordinate gene silencing in mature plants

2019 ◽  
Vol 116 (15) ◽  
pp. 7543-7548 ◽  
Author(s):  
Huan Zhang ◽  
Gozde S. Demirer ◽  
Honglu Zhang ◽  
Tianzheng Ye ◽  
Natalie S. Goh ◽  
...  
Keyword(s):  
Cell Wall ◽  
Gene Silencing ◽  
Plant Cell ◽  
Mammalian Cells ◽  
Plant Cell Wall ◽  
Plant Cells ◽  
Design Parameters ◽  
Dna Nanostructures ◽  
Dna Nanostructure ◽  
The Impact

Delivery of biomolecules to plants relies onAgrobacteriuminfection 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 inNicotiana benthamianaleaves. 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.

scholarly journals Nanotubes effectively deliver siRNA to intact plant cells and protect siRNA against nuclease degradation

2019 ◽  
Author(s):  
Gozde S. Demirer ◽  
Huan Zhang ◽  
Natalie S. Goh ◽  
Roger Chang ◽  
Markita P. Landry
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Sirna Delivery ◽  
Plant Cells ◽  
Plant Biotechnology ◽  
Intact Plant ◽  
Size Exclusion ◽  
Transcriptional Gene Silencing ◽  
Nuclease Degradation

AbstractPost-transcriptional gene silencing (PTGS) is a powerful tool to understand and control plant metabolic pathways, which is central to plant biotechnology. PTGS is commonly accomplished through delivery of small interfering RNA (siRNA) into cells. While siRNA delivery has been optimized for mammalian systems, it remains a significant challenge for plants due to the plant cell wall. Standard plant siRNA delivery methods (Agrobacteriumand viruses) involve coding siRNA into DNA vectors, and are only tractable for certain plant species. Herein, we develop a nanotube-based platform for direct delivery of siRNA, and show high silencing efficiency in intact plant cells. We demonstrate that nanotubes successfully deliver siRNA and silence endogenous genes owing to effective intracellular delivery and nanotube-induced protection of siRNA from nuclease degradation. This study establishes that nanotubes, which are below the size exclusion limit of the plant cell wall, could enable a myriad of plant biotechnology applications that rely on RNA delivery.

Ion Beam Bio-Engineering and Crop Breeding

2010 ◽  
Vol 108-111 ◽  
pp. 536-542
Author(s):  
Tie Gu Wang ◽  
Xue Bin Li ◽  
Tian You Yang ◽  
Wei Dong Wang
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Ion Beam ◽  
Crop Improvement ◽  
Low Energy ◽  
Wall Material ◽  
Crop Breeding ◽  
Exogenous Dna ◽  
Bio Engineering

Ion beam bio-engineering was established by Chinese researcher in 1980s. Over thirty years past, there were delightful progress by low energy ion beam induced mutants and crop improvement. When ion beam implant the organism, charge exchanged between ions and organism surfaces. Meanwhile there were ion sputtering, ion penetration and surface damage of plant cell wall material. Owing to etching action of low energy beam to plant cell wall, ion beam mediated exogenous DNA transformation becomes possible. Representative applications and achievements in crop breeding were described.

scholarly journals DNA Nanostructures Coordinate Gene Silencing in Mature Plants

2019 ◽  
Author(s):  
Huan Zhang ◽  
Gozde S. Demirer ◽  
Honglu Zhang ◽  
Tianzheng Ye ◽  
Natalie S. Goh ◽  
...  
Keyword(s):  
Cell Wall ◽  
Gene Silencing ◽  
Plant Cell ◽  
Mammalian Cells ◽  
Plant Cell Wall ◽  
Plant Cells ◽  
Design Parameters ◽  
Dna Nanostructures ◽  
Dna Nanostructure ◽  
The Impact

AbstractPlant bioengineering may generate high yielding and stress-resistant crops amidst a changing climate and a growing global population (1–3). However, delivery of biomolecules to plants relies onAgrobacteriuminfection (4) or biolistic particle delivery (5), the former of which is only amenable 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 exogenous 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 remains unexplored for passive 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 small interfering RNAs (siRNAs), and effectively silence a constitutively-expressed gene inNicotiana benthamianaleaves. We show that nanostructure internalization into plant cells and the 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 details both the design parameters of importance for plant cell internalization, and also assesses the impact of DNA nanostructure geometry for gene silencing mechanisms.

scholarly journals Plant cell wall biosynthesis: genetic, biochemical and functional genomics approaches to the identification of key genes

2006 ◽  
Vol 4 (2) ◽  
pp. 145-167 ◽  
Author(s):  
Naser Farrokhi ◽  
Rachel A. Burton ◽  
Lynette Brownfield ◽  
Maria Hrmova ◽  
Sarah M. Wilson ◽  
...  
Keyword(s):  
Cell Wall ◽  
Functional Genomics ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Cell Wall Biosynthesis ◽  
Key Genes
Sign in / Sign up

Export Citation Format

Share Document