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Genetic engineering and characterization of Cowpea mosaic virus empty virus-like particles.

Published on Jan 1, 2014in Methods of Molecular Biology
· DOI :10.1007/978-1-62703-751-8_11
Frank Sainsbury21
Estimated H-index: 21
(UQ: University of Queensland),
Pooja Saxena9
Estimated H-index: 9
(JIC: John Innes Centre)
+ 3 AuthorsGeorge P. Lomonossoff52
Estimated H-index: 52
(JIC: John Innes Centre)
Cite
Abstract
The development of methods for the production of empty Cowpea mosaic virus (CPMV) virus-like particles (VLPs) that are devoid of RNA, eVLPs, has renewed promise in CPMV capsid technologies. The recombinant nature of CPMV eVLP production means that the extent and variety of genetic modifications that may be incorporated into the particles is theoretically much greater than those that can be made to infectious CPMV virions due to restrictions on viral propagation of the latter. Free of the infectious agent, the genomic RNA, these particles are now finding potential uses in vaccine development, in vivo imaging, drug delivery, and other nanotechnology applications that make use of internal loading of the empty particles. Here we describe methods for the genetic modification and production of CPMV eVLPs and describe techniques useful for their characterization.
  • References (27)
  • Citations (19)
Cite
References27
Newest
Published on Dec 10, 2012in Biomacromolecules5.67
Amy M. Wen17
Estimated H-index: 17
(Case Western Reserve University),
Sourabh Shukla19
Estimated H-index: 19
(Case Western Reserve University)
+ 8 AuthorsGeorge P. Lomonossoff52
Estimated H-index: 52
(JIC: John Innes Centre)
The development of multifunctional nanoparticles for medical applications is of growing technological interest. A single formulation containing imaging and/or drug moieties that is also capable of preferential uptake in specific cells would greatly enhance diagnostics and treatments. There is growing interest in plant-derived viral nanoparticles (VNPs) and establishing new platform technologies based on these nanoparticles inspired by nature. Cowpea mosaic virus (CPMV) serves as the standard mod...
Published on Mar 12, 2012in Biomacromolecules5.67
Yanhong Wen3
Estimated H-index: 3
,
Lisbeth Grøndahl24
Estimated H-index: 24
+ 3 AuthorsHanne Mørck Nielsen33
Estimated H-index: 33
Dermatan sulfate (DS) is a glycosaminoglycan (GAG) with a great potential as a new therapeutic agent in tissue engineering. The aim of the present study was to investigate the formation of polyelectrolyte complexes (PECs) between chitosan and dermatan sulfate (CS/DS) and delivery of DS from PEC-containing alginate/chitosan/dermatan sulfate (Alg/CS/DS) microspheres for application in tissue regeneration. The CS/DS complexes were initially formed at different conditions including varying CS/DS rat...
Published on Jan 1, 2012in Nanoscale6.97
Alaa A. A. Aljabali13
Estimated H-index: 13
(Norwich Research Park),
J. Elaine Barclay11
Estimated H-index: 11
(Norwich Research Park)
+ 2 AuthorsDavid J. Evans23
Estimated H-index: 23
(Norwich Research Park)
Immobilisation of horseradish peroxidase (HRP) and glucose oxidase (GOX) via covalent attachment of modified enzyme carbohydrate to the exterior of the cowpea mosaic virus (CPMV) capsid gave high retention of enzymatic activity. The number of enzymes bound per virus was determined to be about eleven for HRP and 2–3 for GOX. This illustrates that relatively large biomacromolecules can be readily coupled to the virus surface using simple conjugation strategies. Virus-biomacromolecule hybrids have ...
Published on Nov 4, 2011in ChemBioChem2.59
Frank Sainsbury21
Estimated H-index: 21
(Laval University),
Keith Saunders25
Estimated H-index: 25
(JIC: John Innes Centre)
+ 2 AuthorsGeorge P. Lomonossoff52
Estimated H-index: 52
(JIC: John Innes Centre)
The structure of Cowpea mosaic virus (CPMV) is known to high resolution, thereby enabling the rational use of the particles in diverse applications, from vaccine design to nanotechnology. A recently devised method for the production of empty virus-like particles (eVLPs) has opened up new possibilities for CPMV capsid-based technologies, such as internal mineralisation of the particle. We have investigated the role of the carboxyl (C) terminus of the small coat (S) protein in controlling access t...
Published on Mar 1, 2011in Human Vaccines
Nicholas P. Montague2
Estimated H-index: 2
,
Eva C. Thuenemann9
Estimated H-index: 9
+ 3 AuthorsGeorge P. Lomonossoff52
Estimated H-index: 52
Particles of cowpea mosaic virus (CPMV) have enjoyed considerable success as a means of presenting peptides for vaccine purposes. However, the existing technology has limitations in regard to the size and nature of the peptides which can be presented and has problems regarding bio-containment. Recent developments suggest ways by which these problems can be overcome, increasing the range of potential applications of CPMV-based particle technology.
Published on Jan 1, 2011in Integrative Biology2.77
Alaa A. A. Aljabali13
Estimated H-index: 13
(Norwich Research Park),
Sachin N. Shah4
Estimated H-index: 4
(Norwich Research Park)
+ 2 AuthorsDavid J. Evans23
Estimated H-index: 23
(Norwich Research Park)
The external surface of the plant virus Cowpea mosaic virus (CPMV) can be chemically modified with peptides that direct specific mineralization processes. Subsequent mineralization of the peptide-CPMV conjugates produces monodisperse nanoparticles of ca. 32 nm diameter coated with, for example, cobalt–platinum, iron–platinum or zinc sulfide, which cannot be readily prepared by other methods. This route is particularly attractive as it avoids the need to genetically engineer the protein surface o...
Published on Aug 1, 2010in Nature Protocols11.33
Hon S. Leong17
Estimated H-index: 17
(UWO: University of Western Ontario),
Nicole F. Steinmetz39
Estimated H-index: 39
(Scripps Research Institute)
+ 5 AuthorsJohn D. Lewis23
Estimated H-index: 23
(UWO: University of Western Ontario)
Viral nanoparticles are a novel class of biomolecular agents that take advantage of the natural circulatory and targeting properties of viruses to allow the development of therapeutics, vaccines and imaging tools. We have developed a multivalent nanoparticle platform based on the cowpea mosaic virus (CPMV) that facilitates particle labeling at high density with fluorescent dyes and other functional groups. Compared with other technologies, CPMV-based viral nanoparticles are particularly suited f...
Published on Jul 1, 2010in Annual Review of Phytopathology10.20
Frank Sainsbury21
Estimated H-index: 21
,
M. Carmen Cañizares13
Estimated H-index: 13
,
George P. Lomonossoff52
Estimated H-index: 52
In the 50 years since it was first described, Cowpea mosaic virus (CPMV) has become one of the most intensely studied plant viruses. Research in the past 15 to 20 years has shifted from studying the underlying genetics and structure of the virus to focusing on ways in which it can be exploited in biotechnology. This work led first to the use of virus particles to present peptides, then to the creation of a variety of replicating virus vectors and finally to the development of a highly efficient ...
Published on Apr 9, 2010in Small10.86
Alaa A. A. Aljabali13
Estimated H-index: 13
(Norwich Research Park),
Frank Sainsbury21
Estimated H-index: 21
(Norwich Research Park)
+ 1 AuthorsDavid J. Evans23
Estimated H-index: 23
(Norwich Research Park)
The development of methods for the production of empty Cowpea mosaic virus (CPMV) virus-like particles (VLPs) that are devoid of RNA, eVLPs, has renewed promise in CPMV capsid technologies. The recombinant nature of CPMV eVLP production means that the extent and variety of genetic modifications that may be incorporated into the particles is theoretically much greater than those that can be made to infectious CPMV virions due to restrictions on viral propagation of the latter. Free of the infecti...
Published on Jan 1, 2010in Nanoscale6.97
Alaa A. A. Aljabali13
Estimated H-index: 13
(Norwich Research Park),
J. Elaine Barclay11
Estimated H-index: 11
(Norwich Research Park)
+ 1 AuthorsDavid J. Evans23
Estimated H-index: 23
(Norwich Research Park)
Plant viruses are considered as nanobuilding blocks that can be used as synthons or templates for novel materials. Cowpea mosaic virus (CPMV) particles have been shown to template the fabrication of metallic nanoparticles by an electroless deposition metallization process. Palladium ions were electrostatically bound to the virus capsid and, when reduced, acted as nucleation sites for the subsequent metal deposition from solution. The method, although simple, produced highly monodisperse metallic...
Cited By19
Newest
Published on Jun 6, 2019in Journal of General Virology2.81
Yulia Meshcheriakova5
Estimated H-index: 5
(Norwich Research Park),
George P. Lomonossoff52
Estimated H-index: 52
(Norwich Research Park)
The icosahedral capsid of cowpea mosaic virus is formed by 60 copies of the large (L) and small (S) coat protein subunits. The 24-amino-acid C-terminal peptide of the S coat protein can undergo proteolytic cleavage without affecting particle stability or infectivity. Mutagenic studies have shown that this sequence is involved in particle assembly, virus movement, RNA encapsidation and suppression of gene silencing. However, it is unclear how these processes are related, and which part(s) of the ...
Published on Apr 1, 2019in Advanced Drug Delivery Reviews15.52
Perrin H. Beatty19
Estimated H-index: 19
(U of A: University of Alberta),
John D. Lewis23
Estimated H-index: 23
(U of A: University of Alberta)
Abstract Nanoparticle platforms are particularly attractive for theranostic applications due to their capacity for multifunctionality and multivalency. Some of the most promising nano-scale scaffold systems have been co-opted from nature including plant viruses such as cowpea mosaic virus (CPMV). The use of plant viruses like CPMV as viral nanoparticles is advantageous for many reasons; they are non-infectious and nontoxic to humans and safe for use in intravital imaging and drug delivery. The C...
Published on Feb 1, 2019in Biochimie3.36
Jaison Jeevanandam5
Estimated H-index: 5
(Curtin University),
Kaushik Pal10
Estimated H-index: 10
(Bharath University)
+ 0 AuthorsMichael K. Danquah1
Estimated H-index: 1
(UTC: University of Tennessee at Chattanooga)
Abstract Viruses are considered as natural nanomaterials as they are in the size range of 20–500 nm with a genetical material either DNA or RNA, which is surrounded by a protein coat capsid. Recently, the field of virus nanotechnology is gaining significant attention from researchers. Attention is given to the utilization of viruses as nanomaterials for medical, biotechnology and energy applications. Removal of genetic material from the viral capsid creates empty capsid for drug incorporation an...
Published on Oct 24, 2018in Journal of Virology4.32
Inga Kruse3
Estimated H-index: 3
(JIC: John Innes Centre),
Hadrien Peyret6
Estimated H-index: 6
(JIC: John Innes Centre)
+ 1 AuthorsGeorge P. Lomonossoff52
Estimated H-index: 52
(JIC: John Innes Centre)
ABSTRACT To elucidate the linkage between replication and encapsidation in Picornavirales , we have taken advantage of the bipartite nature of a plant-infecting member of this order, cowpea mosaic virus (CPMV), to decouple the two processes. RNA-free virus-like particles (empty virus-like particles [eVLPs]) can be generated by transiently coexpressing the RNA-2-encoded coat protein precursor (VP60) with the RNA-1-encoded 24,000-molecular-weight (24K) protease, in the absence of the replication m...
Published on Jan 1, 2018in Methods of Molecular Biology
Christina Dickmeis4
Estimated H-index: 4
(RWTH Aachen University),
Klara Altintoprak3
Estimated H-index: 3
(University of Stuttgart)
+ 2 AuthorsUlrich Commandeur25
Estimated H-index: 25
(RWTH Aachen University)
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 an...
Published on Jan 1, 2018
Carissa M. Soto20
Estimated H-index: 20
(NRL: United States Naval Research Laboratory),
Walter J. Dressick30
Estimated H-index: 30
(NRL: United States Naval Research Laboratory)
Published on Jan 1, 2018
Roger Castells-Graells1
Estimated H-index: 1
(Norwich Research Park),
George P. Lomonossoff52
Estimated H-index: 52
(Norwich Research Park),
Keith Saunders25
Estimated H-index: 25
(Norwich Research Park)
Published on Jan 1, 2018
Alaa A. A. Aljabali13
Estimated H-index: 13
(YU: Yarmouk University),
David J. Evans23
Estimated H-index: 23
(Norwich Research Park)
Published on Dec 15, 2017in Biochemical Society Transactions4.29
Yulia Meshcheriakova5
Estimated H-index: 5
(Norwich Research Park),
Alex Durrant1
Estimated H-index: 1
(Norwich Research Park)
+ 2 AuthorsGeorge P. Lomonossoff52
Estimated H-index: 52
(Norwich Research Park)
Particles of cowpea mosaic virus (CPMV) have enjoyed considerable success as nanoparticles. The development of a system for producing empty virus-like particles (eVLPs) of the virus, which are non-infectious and have the potential to be loaded with heterologous material, has increased the number of possible applications for CPMV-based particles. However, for this potential to be realised, it was essential to demonstrate that eVLPs were accurate surrogates for natural virus particles, and this in...
Published on Aug 18, 2017in Frontiers in Plant Science4.11
Keith Saunders25
Estimated H-index: 25
(Norwich Research Park),
George P. Lomonossoff52
Estimated H-index: 52
(Norwich Research Park)
We have utilised plant-based transient expression to produce tobacco mosaic virus (TMV)-based nano-rods of predetermined lengths. This is achieved by expressing RNAs containing the TMV origin of assembly sequence (OAS) and the sequence of the TMV coat protein either on the same RNA molecule or on two separate constructs. We show that the length of the resulting nano-rods is dependent upon the length of the RNA that possesses the OAS element. By expressing a version of the TMV coat protein that i...