It is widely accepted that significant differences between the in-vivo virus cycle and its in-vitro representations exist. In-vitro assays lack the cellular regulatory mechanisms that influence the pathways of viral protein synthesis and self-assembly. We propose to implement a demonstration of a new in-vivo imaging technology, based on virus-like particles (VLPs) and able to bridge the existing gap between in-vitro and in-vivo experiments. Virus-like particles are here hybrid biological/inorganic complexes composed of a viral capsid encapsulating a functionalized nanoparticle instead of nucleic acid. The proposed in vivo imaging technology is of potential high impact since it is not limited to virus assembly but it can be extended to many other self-organizing macromolecular complexes occurring in the confines of a cell. An illustration of the new capabilities is provided through the following proof-of-concept experiments: a) determine real-time disassembly trajectories of individual VLPs in vitro and in vivo with 10 nm spatial resolution and 10 ms time resolution;b) locate individual self-assembled VLPs in cells and their spatial relation with the sites for viral replication. The anticipated results from this proposal will provide a new non-intrusive imaging technology coupled to a means of measuring in-vivo sub-cellular dynamics and will teach us about the rules for virus particle assembly, disassembly, and intracellular transit. We will learn unprecedented basic insight into virus biology. The technology will also broaden the area of future VLP applications and yield a new class of high-output probes that can be adapted to a wide range of targets.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM081029-05
Application #
8144249
Study Section
Special Emphasis Panel (ZRG1-BST-R (51))
Program Officer
Deatherage, James F
Project Start
2007-09-01
Project End
2013-02-28
Budget Start
2011-09-01
Budget End
2013-02-28
Support Year
5
Fiscal Year
2011
Total Cost
$266,601
Indirect Cost
Name
Indiana University Bloomington
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
006046700
City
Bloomington
State
IN
Country
United States
Zip Code
47401
Cheng, Fan; Tsvetkova, Irina B; Khuong, Y-Lan et al. (2013) The packaging of different cargo into enveloped viral nanoparticles. Mol Pharm 10:51-8
Tsvetkova, Irina B; Cheng, Fan; Ma, Xiang et al. (2013) Fusion of mApple and Venus fluorescent proteins to the Sindbis virus E2 protein leads to different cell-binding properties. Virus Res 177:138-46
Yildiz, Ibrahim; Tsvetkova, Irina; Wen, Amy M et al. (2012) Engineering of Brome mosaic virus for biomedical applications. RSC Adv 2:3670-3677
Vieweger, Mario; Goicochea, Nancy; Koh, Eun Sohl et al. (2011) Photothermal imaging and measurement of protein shell stoichiometry of single HIV-1 Gag virus-like nanoparticles. ACS Nano 5:7324-33
Huang, Xinlei; Stein, Barry D; Cheng, Hu et al. (2011) Magnetic virus-like nanoparticles in N. benthamiana plants: a new paradigm for environmental and agronomic biotechnological research. ACS Nano 5:4037-45
Goicochea, Nancy L; Datta, Siddhartha A K; Ayaluru, Murali et al. (2011) Structure and stoichiometry of template-directed recombinant HIV-1 Gag particles. J Mol Biol 410:667-80
Vieweger, Mario; Jiang, Xuan; Lim, Young-Kwan et al. (2011) Conformationally dynamic ?-conjugation: probing structure-property relationships of fluorescent tris(N-salicylideneaniline)s. J Phys Chem A 115:13298-308
Bronstein, Lyudmila M; Shtykova, Eleonora V; Malyutin, Andrey et al. (2010) Hydrophilization of Magnetic Nanoparticles with Modified Alternating Copolymers. Part 1: The Influence of the Grafting. J Phys Chem C Nanomater Interfaces 114:21900-21907
DuFort, Christopher C; Dragnea, Bogdan (2010) Bio-enabled synthesis of metamaterials. Annu Rev Phys Chem 61:323-44
Daniel, Marie-Christine; Tsvetkova, Irina B; Quinkert, Zachary T et al. (2010) Role of surface charge density in nanoparticle-templated assembly of bromovirus protein cages. ACS Nano 4:3853-60

Showing the most recent 10 out of 18 publications