Construction of gold-siRNANPR1 nanoparticles for effective and quick silencing of NPR1 in Arabidopsis thaliana

Lei, Wenxue; An, Zi-Shuai; Zhang, Baihong; Wu, Qian; Gong, Wenrong; Li, Jinming; Chen, Wenli

doi:10.1039/d0ra02156c

Cited by 33 publications

(26 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…After preparation of PDMAEMA + DNA and PEI + DNA complexes containing 10 μg DNA at a N/P ratio of 15 (total volume 10 μL), it was diluted with 10 mM MgCl 2 /MES with pH 5.7 to a final volume of 200 μL and then infiltrated on the abaxial surface of the leaves using a 1-mL needleless syringe by applying gentle pressure. FAM-siRNA NPR1 (20 μM) ( Lei et al, 2020 ) was mixed with PDMAEMA (1 mg/mL) at the fixed N/P ratio of 15 and incubated at room temperature for 30 min. The formed complexes were diluted 10 times in 10 mM MgCl 2 /MES (pH 5.7) before infiltration.…”

Section: Methodsmentioning

confidence: 99%

“…In our previous study, functioned gold nanoparticles were used to carry a small interfering RNA and successfully silenced a target gene ( NPR1 ) in Arabidopsis thaliana ( Lei et al, 2020 ). To further explore the transfection behavior of PDMAEMA in plant cells, we combine plasmid GFP (pGFP) as a reporter gene with PDMAEMA and incubated them with protoplasts and leaves to transfect plant cells.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Efficient Transient Expression of Plasmid DNA Using Poly (2-(N,N-Dimethylamino) Ethyl Methacrylate) in Plant Cells

Cao

Zhang

et al. 2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Nanomaterials have been widely studied for their potential to become the new generation of nanocarriers in gene transfection, yet it remains still difficult to apply them efficiently and succinctly to plant cells. Poly (2-(N,N-dimethylamino) ethyl methacrylate) (PDMAEMA), which possesses temperature and pH dual-sensitivity, has largely been applied in animal cells, but it is rarely involved in plant cells. As a proof of concept, PDMAEMA as a gene carrier is incubated with plasmid GFP (pGFP) to explore its transfection ability in plants, and cationic polymer polyethylenimine (PEI) is used as a control. pGFP was efficiently condensed into the nanostructure by electrostatic interactions at an N/P (amino group from cationic polymers/phosphate group from plasmid DNA (pDNA)) ratio of 15; after complexation into nanocarriers, pGFP was protected from endonuclease degradation according to the DNase I digestion assay. After incubation with protoplasts and leaves, GFP was observed with confocal microscopy in plant cells. Western blot experiments confirmed GFP expression at the protein level. Toxicity assay showed PDMAEMA had a lower toxicity than PEI. These results showed that transient expression of pGFP was readily achieved in Arabidopsis thaliana and Nicotiana benthamiana. Notably, PDMAEMA showed lower cytotoxicity than PEI upon incubation with Nicotiana benthamiana leaves. PDMAEMA exhibited great potency for DNA delivery in plant cells. This work provides us with new ideas of more concise and more effective methods for plant transformation.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Efficient Transient Expression of Plasmid DNA Using Poly (2-(N,N-Dimethylamino) Ethyl Methacrylate) in Plant Cells

Cao

Zhang

et al. 2022

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…Our previous reports revealed that AuNPs-PEI-FAM-siRNA NPR1 successfully silenced the function of NPR1 ( Lei et al., 2020 ). Here, we infiltrated healthy periwinkle leaves with AuNPs-PEI-FAM-siRNA NPR1 and used the fluorescence of FAM-siRNA NPR1 to follow the entry of AuNPs-PEI into the nucleus.…”

Section: Resultsmentioning

confidence: 99%

“…It has been previously confirmed that AuNPs-PEI carrying exogenous siRNA NPR1 successfully silenced the NPR1 gene with high specificity ( Lei et al., 2020 ). However, the timing of AuNPs-PEI entry into the cytoplasm and nucleus has not been further investigated.…”

Section: Introductionmentioning

confidence: 89%

Expression of PEI-coated gold nanoparticles carrying exogenous gene in periwinkle mesophyll cells and its practice in huanglongbing research

et al. 2022

Self Cite

View full text Add to dashboard Cite

“…Nanocomposites not only improve the utilization of drugs, they reduce the usage of drugs and also alleviate environmental pressure. Because we successfully silenced the NPR1 gene using AuNPs carrying siRNA NPR1 ( Lei et al, 2020 ). Our laboratory is developing a nano-genetic technology to activate a defense system for improving resistance by transferring resistant genes to hosts through nanomaterials, hoping to contribute to the prevention and treatment of HLB.…”

Section: Chemical Methodsmentioning

confidence: 99%

Controlling Citrus Huanglongbing: Green Sustainable Development Route Is the Future

Ruan

Zhou

et al. 2021

Front. Plant Sci.

View full text Add to dashboard Cite

Huanglongbing (HLB) is the most severe bacterial disease of citrus crops caused by Candidatus Liberibacter spp. It causes a reduction in fruit yield, poor fruit quality, and even plants death. Due to the lack of effective medicine, HLB is also called citrus “AIDS.” Currently, it is essential for the prevention and control of HLB to use antibiotics and pesticides while reducing the spread of HLB by cultivating pathogen-free seedlings, removing disease trees, and killing Asian citrus psyllid (ACP). New compounds [e.g., antimicrobial peptides (AMPs) and nanoemulsions] with higher effectiveness and less toxicity were also found and they have made significant achievements. However, further evaluation is required before these new antimicrobial agents can be used commercially. In this review, we mainly introduced the current strategies from the aspects of physical, chemical, and biological and discussed their environmental impacts. We also proposed a green and ecological strategy for controlling HLB basing on the existing methods and previous research results.

show abstract

Construction of gold-siRNA_NPR1 nanoparticles for effective and quick silencing of NPR1 in Arabidopsis thaliana

Abstract: Gold nanoparticles (AuNPs) have been widely used as gene silencing agents and therapeutics for treatment due to their high transfection efficiency and lack of cytotoxicity, but their roles in gene silencing in plants have not yet been reported.

Cited by 33 publications

References 41 publications

Efficient Transient Expression of Plasmid DNA Using Poly (2-(N,N-Dimethylamino) Ethyl Methacrylate) in Plant Cells

Efficient Transient Expression of Plasmid DNA Using Poly (2-(N,N-Dimethylamino) Ethyl Methacrylate) in Plant Cells

Expression of PEI-coated gold nanoparticles carrying exogenous gene in periwinkle mesophyll cells and its practice in huanglongbing research

Controlling Citrus Huanglongbing: Green Sustainable Development Route Is the Future

Contact Info

Product

Resources

About