scholarly journals In vitro-reassembled plant virus-like particles for loading of polyacids

2006 ◽  
Vol 87 (9) ◽  
pp. 2749-2754 ◽  
Author(s):  
Yupeng Ren ◽  
Sek-Man Wong ◽  
Lee-Yong Lim
Keyword(s):  
Electrostatic Interactions ◽  
Plant Viruses ◽  
Virus Like Particles ◽  
Protein Cages ◽  
Large Molecules ◽  
Protein Cage ◽  
Drug Molecules ◽  
Structural Conformation ◽  
Foreign Materials

The coat protein (CP) of certain plant viruses may reassemble into empty virus-like particles (VLPs) and these protein cages may serve as potential drug delivery platforms. In this paper, the production of novel VLPs from the Hibiscus chlorotic ringspot virus (HCRSV) is reported and the capacity to load foreign materials was characterized. VLPs were readily produced by destabilizing the HCRSV in 8 M urea or Tris buffer pH 8, in the absence of calcium ions, followed by removal of viral RNA by ultrahigh-speed centrifugation and the reassembly of the CP in sodium acetate buffer pH 5. The loading of foreign materials into the VLPs was dependent on electrostatic interactions. Anionic polyacids, such as polystyrenesulfonic acid and polyacrylic acid, were successfully loaded but neutrally charged dextran molecules were not. The molecular-mass threshold for the polyacid cargo was about 13 kDa, due to the poor retention of smaller molecules, which readily diffused through the holes between the S domains present on the surface of the VLPs. These holes precluded the entry of large molecules, but allowed smaller molecules to enter or exit. The polyacid-loaded VLPs had comparable size, morphology and surface-charge density to the native HCRSV, and the amount of polyacids loaded was comparable to the weight of the native genomic materials. The conditions applied to disassembly–reassembly of the virions did not change the structural conformation of the CP. HCRSV-derived VLPs may provide a promising nano-sized protein cage for delivery of anionic drug molecules.

scholarly journals Encapsidation of DNA, a protein and a fluorophore into virus-like particles by the capsid protein of cucumber mosaic virus

2012 ◽  
Vol 93 (5) ◽  
pp. 1120-1126 ◽  
Author(s):  
Xiaoyun Lu ◽  
Jeremy R. Thompson ◽  
Keith L. Perry
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Capsid Protein ◽  
Plant Viruses ◽  
Insect Vector ◽  
Virus Like Particles ◽  
Protein Cage ◽  
Transmission Electron ◽  
Lower Size

An important property of some spherical plant viruses is their ability to reassemble in vitro from native capsid protein (CP) and RNA into infectious virus-like particles (VLPs). Virions of cucumber mosaic virus (CMV) are stabilized by protein–RNA interactions and the nucleic acid is essential for assembly. This study demonstrated that VLPs will form in the presence of both ssDNA and dsDNA oligonucleotides, and with a lower size limit of 20 nt. Based on urea disruption assays, assembled VLPs from CMV CP and RNA (termed ReCMV) exhibited a level of stability similar to that of virions purified from plants, whilst VLPs from CMV CP and a 20mer exhibited comparable or greater stability. Fluorescent labelling of VLPs was achieved by the encapsidation of an Alexa Fluor 488-labelled 45mer oligonucleotide (ReCMV-Alexa488-45) and confirmed by transmission electron and confocal microscopy. Using ssDNA as a nucleating factor, encapsidation of fluorescently labelled streptavidin (53 kDa) conjugated to a biotinylated oligonucleotide was observed. The biological activity and stability of ReCMV and ReCMV-Alexa488-45 was confirmed in infectivity assays and insect vector feeding assays. This work demonstrates the utility of CMV CP as a protein cage for use in the growing repertoire of nanotechnological applications.

Synthesis of Metal Core in the Apoferritin Cavity

2012 ◽  
Vol 549 ◽  
pp. 216-220
Author(s):  
Ting Wang
Keyword(s):  
Particle Size ◽  
Electrostatic Interactions ◽  
Molecular Level ◽  
Reaction Vessel ◽  
Inorganic Materials ◽  
Metal Core ◽  
Protein Cages ◽  
Protein Cage ◽  
Protein Cavity ◽  
Uniform Particle Size

Many methods to produce NPs have been studied. Among them, the molecular level understanding of the formation of solids in biological systems has provided inspiration for the controlled formation of novel inorganic materials. The use of a protein cavity as the growth field of NPs provides one candidate method for obtaining uniform particle size. The host-guest relationship between these protein cages and the encapsulated material is based primarily on a complementary electrostatic interaction. Charged interfaces play important roles in defining electrostatically distinct environments for spatially defined encapsulation. The electrostatic interactions can be approximated by Guoy-Chapman theory of charged interfaces. These interactions are proposed at the interior of protein cages such as ferritin and viruses. The protein cage with a highly charged interior surface with pores that allow molecular access to the inside of the protein could act as a constrained reaction vessel. The present paper will focus on the synthesis of Metal NPs using ferritin.

scholarly journals Polyelectrolyte Encapsulation and Confinement within Protein Cage-Inspired Nanocompartments

Pharmaceutics ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1551
Author(s):  
Qing Liu ◽  
Ahmed Shaukat ◽  
Daniella Kyllönen ◽  
Mauri A. Kostiainen
Keyword(s):  
Small Molecules ◽  
Driving Force ◽  
Electrostatic Interactions ◽  
Inorganic Nanoparticles ◽  
Wild Type ◽  
Potential Toxicity ◽  
Protein Cages ◽  
Functional Sites ◽  
Protein Cage ◽  
Or Gene

Protein cages are nanocompartments with a well-defined structure and monodisperse size. They are composed of several individual subunits and can be categorized as viral and non-viral protein cages. Native viral cages often exhibit a cationic interior, which binds the anionic nucleic acid genome through electrostatic interactions leading to efficient encapsulation. Non-viral cages can carry various cargo, ranging from small molecules to inorganic nanoparticles. Both cage types can be functionalized at targeted locations through genetic engineering or chemical modification to entrap materials through interactions that are inaccessible to wild-type cages. Moreover, the limited number of constitutional subunits ease the modification efforts, because a single modification on the subunit can lead to multiple functional sites on the cage surface. Increasing efforts have also been dedicated to the assembly of protein cage-mimicking structures or templated protein coatings. This review focuses on native and modified protein cages that have been used to encapsulate and package polyelectrolyte cargos and on the electrostatic interactions that are the driving force for the assembly of such structures. Selective encapsulation can protect the payload from the surroundings, shield the potential toxicity or even enhance the intended performance of the payload, which is appealing in drug or gene delivery and imaging.

scholarly journals Innovative Applications of Plant Viruses in Drug Targeting and Molecular Imaging- A review

2020 ◽  
Vol 16 ◽  
Author(s):  
Alaa A A Aljabali ◽  
Mazhar S. Al Zoubi ◽  
Khalid M. Al-Batayneh ◽  
Dinesh M. Pardhi ◽  
Kamal Dua ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Molecular Imaging ◽  
Plant Viruses ◽  
Antimicrobial Drugs ◽  
Protein Cages ◽  
Specific Subset ◽  
Potential Applications ◽  
Viral Nanoparticles

Background: Nature had already engineered various types of nanoparticles (NPs), especially viruses, which can deliver their cargo to the host/targeted cells. The ability to selectively target specific cells offers a significant advantage over the conventional approach. Numerous organic NPs, including native protein cages, virus-like pieces, polymeric saccharides, and liposomes, have been used for the preparation of nanoparticulate. Such nanomaterials have demonstrated better performance and as well as improved biocompatible, devoid of side effects, and stable without any deterioration. Objective: This review discusses current clinical and scientific research on naturally occurring nanomaterials. The review illustrates and updates the tailor-made approaches for selective delivery and targeted medications that require a highaffinity interconnection to the targeted cells. Method: A comprehensive search was performed using keywords for viral nanoparticles, viral particles for drug delivery, viral nanoparticles for molecular imaging, theranostics applications of viral nanoparticles and plant viruses in nanomedicine. We searched in Google Scholar, PubMed, Springer, Medline, and Elsevier from 2000 to till date and by the bibliographic review of all identified articles. Results: The findings demonstrated that structures dependent on nanomaterials might have potential applications in diagnostics, cell marking, comparing agents (computed tomography and magnetic resonance imaging), and antimicrobial drugs, as well as drug delivery structures. However, measures should be taken in order to prevent or mitigate in pharmaceutical or medical applications the toxic impact or incompatibility of nanoparticle-based structures with biological systems. Conclusion: The review provided an overview of the latest advances in nanotechnology, outlining the difficulties and the advantages of in vivo and in vitro structures that are focused on a specific subset of the natural nanomaterials.

Characteristics of Artificial Virus-like Particles Assembled in vitro from Potato Virus X Coat Protein and Foreign Viral RNAs

Acta Naturae ◽  
2011 ◽  
Vol 3 (3) ◽  
pp. 40-46 ◽  
Author(s):  
M V Arkhipenko ◽  
E K Petrova ◽  
N A Nikitin ◽  
A D Protopopova ◽  
E V Dubrovin ◽  
...  
Keyword(s):  
Coat Protein ◽  
Potato Virus ◽  
Potato Virus X ◽  
Viral Rnas ◽  
Virus Like Particles ◽  
Artificial Virus

Programmable in vitro Co-Encapsidation of Guest Proteins Inside Protein Nanocages

2018 ◽  
Author(s):  
Noor H. Dashti ◽  
Rufika S. Abidin ◽  
Frank Sainsbury
Keyword(s):  
Self Assembly ◽  
Mammalian Cells ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Super Resolution ◽  
Cell Engineering ◽  
Particle Analysis ◽  
Protein Cages

Bioinspired self-sorting and self-assembling systems using engineered versions of natural protein cages have been developed for biocatalysis and therapeutic delivery. The packaging and intracellular delivery of guest proteins is of particular interest for both <i>in vitro</i> and <i>in vivo</i> cell engineering. However, there is a lack of platforms in bionanotechnology that combine programmable guest protein encapsidation with efficient intracellular uptake. We report a minimal peptide anchor for <i>in vivo</i> self-sorting of cargo-linked capsomeres of the Murine polyomavirus (MPyV) major coat protein that enables controlled encapsidation of guest proteins by <i>in vitro</i> self-assembly. Using Förster resonance energy transfer (FRET) we demonstrate the flexibility in this system to support co-encapsidation of multiple proteins. Complementing these ensemble measurements with single particle analysis by super-resolution microscopy shows that the stochastic nature of co-encapsidation is an overriding principle. This has implications for the design and deployment of both native and engineered self-sorting encapsulation systems and for the assembly of infectious virions. Taking advantage of the encoded affinity for sialic acids ubiquitously displayed on the surface of mammalian cells, we demonstrate the ability of self-assembled MPyV virus-like particles to mediate efficient delivery of guest proteins to the cytosol of primary human cells. This platform for programmable co-encapsidation and efficient cytosolic delivery of complementary biomolecules therefore has enormous potential in cell engineering.

Synthesis, Characterization, Antimicrobial and Antioxidant Potential of Diazenyl Chalcones

2018 ◽  
Vol 18 (10) ◽  
pp. 844-856 ◽  
Author(s):  
Harmeet Kaur ◽  
Balasubramanian Narasimhan
Keyword(s):  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Antioxidant Potential ◽  
Dilution Method ◽  
Dpph Assay ◽  
Antimicrobial Potential ◽  
Structural Conformation ◽  
Uv Visible ◽  
Tube Dilution Method

A series of diazenyl chalcones was prepared by base catalyzed Claisen-Schmidt condensation of synthesized hydroxy substituted acetophenone azo dye with various substituted aromatic/ heteroaromatic aldehydes. The structural conformation of synthesized chalcones was done by a number of physicochemical and spectral means like FTIR, UV-visible, mass, NMR spectroscopy and CHNS/O analysis. These diazenyl chalcones were assessed for their in vitro antimicrobial potential against several Gram-negative, Gram-positive bacterial and fungal strains by serial tube dilution method. The fluconazole and cefadroxil were used as standard drugs. The target compounds were also evaluated for their antioxidant potential by DPPH assay. (2E)-3-(2,4-Dichlorophenyl)-1-(4-((2,6- dihydroxyphenyl)diazenyl)phenyl)prop-2-en-1-one (C-7) had shown very good antimicrobial potential with MIC ranges from 3.79 to 15.76 μg/ml against most of the tested microorganisms. Most of the synthesized diazenyl chalcones were found to be active against B. subtilis. The (2E)-1-(5-((2-Chloro- 4-nitrophenyl)diazenyl)-2-hydroxyphenyl)-3-(2-hydroxynaphthalen-1-yl)prop-2-en-1-one (C-10) had shown high free radical-scavenging activity when compared with the ascorbic acid as the reference antioxidant.

a Novel Combinational Nanodrug Delivery System Induces Synergistic Inhibition of Lung Adenocarcinoma Cells in vitro

2020 ◽  
Vol 17 ◽  
Author(s):  
Mingliang Fan ◽  
Jiping Li
Keyword(s):  
Lung Adenocarcinoma ◽  
Delivery System ◽  
The Body ◽  
Anticancer Efficacy ◽  
Nanodrug Delivery ◽  
Drug Molecules ◽  
Cellular Assays ◽  
Lung Adenocarcinoma Cell Line ◽  
M Phase

Background: The combination of two or more therapeutic drugs is an attractive approach to improve the treatment of experimental tumors. Leveraging nanocarriers for combinational drug delivery can allow a control over drug biological fate and promote co-localization in the same area of the body. However, there are certain concerns regarding the biodegradability and potential long-term toxicity arising from these synthetic nanoscale carriers. Objective: Our aim was to develop a combinational nanodrug delivery system formed by self-assembling of amphiphilic drug molecules，minimizing potential toxicities associated with using additional synthetic nanocarriers. Methods: A novel prodrug chlorambucil gemcitabine conjugate was synthesized, this prodrug was used for the encapsulation of an additional hydrophobic anticancer drug paclitaxel, taking the form of combinational nanodrugs. Particle size and zeta potential were evaluated, cytotoxicity assay and apoptosis/cell cycle analysis were also performed to validate the anticancer efficacy of the combinational nanodrugs. Results: The combinational nanodrugs were acquired by means of nanoprecipitation. In A549 lung adenocarcinoma cell line, cellular assays revealed that co-delivery of low dosage paclitaxel with chlorambucil gemcitabine conjugate can act synergistically to inhibit cell growth and induce accumulation of cells in the G2/M phase with a concomitant decrease in G0/G1 compartment. Conclusion: Chlorambucil gemcitabine conjugate and paclitaxel can co-assemble into composite nanoparticles by a nanoprecipitation process and the resulting combinational nanodrugs showed synergistic anticancer effect. This synthetic nanocarrier-free approach might broaden the nanodrug concept and have potential in cancer therapy.

scholarly journals Towards plant resistance to viruses using protein-only RNase P

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Anthony Gobert ◽  
Yifat Quan ◽  
Mathilde Arrivé ◽  
Florent Waltz ◽  
Nathalie Da Silva ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Virus Replication ◽  
Yield Loss ◽  
Rnase P ◽  
Plant Viruses ◽  
Yellow Mosaic Virus ◽  
Resistance To Viruses ◽  
Target Plant

AbstractPlant viruses cause massive crop yield loss worldwide. Most plant viruses are RNA viruses, many of which contain a functional tRNA-like structure. RNase P has the enzymatic activity to catalyze the 5′ maturation of precursor tRNAs. It is also able to cleave tRNA-like structures. However, RNase P enzymes only accumulate in the nucleus, mitochondria, and chloroplasts rather than cytosol where virus replication takes place. Here, we report a biotechnology strategy based on the re-localization of plant protein-only RNase P to the cytosol (CytoRP) to target plant viruses tRNA-like structures and thus hamper virus replication. We demonstrate the cytosol localization of protein-only RNase P in Arabidopsis protoplasts. In addition, we provide in vitro evidences for CytoRP to cleave turnip yellow mosaic virus and oilseed rape mosaic virus. However, we observe varied in vivo results. The possible reasons have been discussed. Overall, the results provided here show the potential of using CytoRP for combating some plant viral diseases.

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