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Plant virus capsids are attractive entities for nanotechnological applications because of their variation in shape and natural assembly ability. This chapter describes the production and modification of three differently shaped plant virus capsids for silica mineralization purposes. The chosen plant viruses exhibit either an icosahedral (cowpea mosaic virus, CPMV), or a flexuous rod-like structure (potato virus X, PVX), or a rigid rod-like shape (tobacco mosaic virus, TMV), and are well-known and frequently used plant viruses for biotechnological applications. We describe the production (including genetic or chemical modification) and purification of the plant viruses or of empty virus-like particles in the case of CPMV, as well as the characterization of these harvested templates. The mineralization procedures and differences in the protocols specific to the distinct viruses are described, and the analyses of the mineralization results are explained.

Due to the nanoscale size and the strictly controlled and consistent morphologies of viruses, there has been a recent interest in utilizing them in nanotechnology. The structure, surface chemistries and physical properties of many viruses have been well elucidated, which have allowed identification of regions of their capsids which can be modified either chemically or genetically for nanotechnological uses. In this review we focus on the use of such modifications for the functionalization and production of viruses and empty viral capsids that can be readily decorated with metals in a highly tuned manner. In particular, we discuss the use of two plant viruses (Cowpea mosaic virus and Tobacco mosaic virus) which have been extensively used for production of novel metal nanoparticles (<100nm), composites and building blocks for 2D and 3D materials, and illustrate their applications., (© 2013 Elsevier Inc. All rights reserved.)

Chemical addressability of viral particles has played a pivotal role in adapting these biogenic macromolecules for various applications ranging from medicine to inorganic catalysis. Cowpea mosaic virus possesses multiple features that are advantageous for the next generation of virus-based nanotechnology: consistent multimeric assemblies dictated by its genetic code, facile large scale production, and lack of observable toxicity in humans. Herein, the chemistry of the viral particles is extended with the use of Cu-free strain-promoted azide-alkyne cycloaddition reaction, or SPAAC reaction. The elimination of Cu, its cocatalyst and reducing agent, simplifies the reaction scheme to a more straightforward approach, which can be directly applied to living systems. As a proof of concept, the viral particles modified with the azadibenzylcyclooctyne functional groups are utilized to trigger and amplify a weak fluorescent signal (azidocoumarin) in live cell cultures to visualize the non-natural sugars. Future adaptations of this platform may be developed to enhance biosensing applications.

Surface engineering of plant virus capsids via cationization (1) and stoichiometric coupling of a polymer surfactant coronal layer (2) produces a highly concentrated, solvent-free liquid virus at 28 °C. These ionic bionanoconstructs are viscoelastic, retain plant infectivity and can be dispersed in a range of organic solvents for aerosol delivery., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A quintessential tenet of nanotechnology is the self-assembly of nanometer-sized components into devices. Biological macromolecular systems such as viral particles were found to be suitable building blocks for nanotechnology for several reasons: viral capsids are extremely robust and can be produced in large quantities with ease, the particles self-assemble into monodisperse particles with a high degree of symmetry and polyvalency, they have the propensity to form arrays, and they offer programmability through genetic and chemical engineering. Here, we review the recent advances in engineering the icosahedral plant virus Cowpea mosaic virus (CPMV) for applications in nano-medicine and -technology. In the first part, we will discuss how the combined knowledge of the structure of CPMV at atomic resolution and the use of chimeric virus technology led to the generation of CPMV particles with short antigenic peptides for potential use as vaccine candidates. The second part focuses on the chemical addressability of CPMV. Strategies to chemically attach functional molecules at designed positions on the exterior surface of the viral particle are described. Biochemical conjugation methods led to the fabrication of electronically conducting CPMV particles and networks. In addition, functional proteins for targeted delivery to mammalian cells were successfully attached to CPMV. In the third part, we focus on the utilization of CPMV as a building block for the generation of 2D and 3D arrays. Overall, the potential applications of viral nanobuilding blocks are manifold and range from nanoelectronics to biomedical applications.

We have developed a substrate-based bottom-up approach to assemble two different color emitting quantum dots (CdSe/ZnS core/shell QDs) on the surface of a novel virus mutant, CPMV-T184C. Electrical characteristics of individual hybrids were investigated by conductive atomic force microscopy for potential digital memory applications (i.e., RAM). These individual 40 nm CPMV-QD(1,2) hybrids exhibited reversible bistable electrical behavior during repeatable writing-reading-erasing processes at the nanoscale.

Virus-like particles are biomimetic delivery vehicles that cloak nanoscale cores inside coatings of viral capsid proteins, offering the potential for protecting their contents and targeting them to particular tissues and cells. To date, encapsidation has been demonstrated only for a relatively limited variety of core materials, such as compressible polymers and facetted nanocrystals, over a narrow range of cores sizes and of pH and ionic strength. Here, we encapsidate spherical nanodroplets of incompressible oil stabilized by adsorbed anionic surfactant using cationic capsid protein purified from cowpea chlorotic mottle virus. By imaging with transmission electron microscopy we show that, as the droplets become larger than the wild-type RNA core, the protein is forced to self-assemble into spherical shells that are not perfect icosahedra having special triangulation numbers characteristic of the Caspar-Klug hierarchy. Consequently, the distribution of protein conformations on larger droplets is significantly different than in the wild-type shell.

The generation of long-range three-dimensional nanoscopic patterns is a major goal in materials chemistry. Here we report a strategy for creating such systems using virus crystals as scaffolds which can be infiltrated with metal specifically palladium and platinum. The inorganic component effectively packs within the porous macromolecular crystal architecture, providing a route for patterning these materials on the nanometer length scale. To verify the quality of the metal infiltration, SEM-EDX was used to determine the homogeneous distribution of metal across the crystal, and TEM was used to confirm that the metal was confined within the porous structure of the crystal.

We have developed a multistep route to the fabrication of virus assembled nanostructures with chemoselective protein-to-surface linkers synthesized by an efficient solid-phase method. These linkers were used to create patterns of 30-to-50-nm-width-lines by scanning probe nanolithography. Genetically modified cow pea mosaic virus with unique cysteine residues at specific locations on their capsomers were assembled through covalent linkage on these patterns. The morphology of the assembled structures on these line patterns characterized by atomic force microscopy was found to be strongly influenced by the intervirion interactions.