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Polymorphic assembly of virus-capsid proteins around DNA and the cellular uptake of the resulting particles
- Source :
- Journal of controlled release, 307, 342-354. Elsevier
- Publication Year :
- 2019
-
Abstract
- Virus-like particles (VLPs), i.e. molecular assemblies that resemble the geometry and organization of viruses, are promising platforms for therapeutics and imaging. Understanding the assembly and cellular uptake pathways of VLPs can contribute to the development of new antiviral drugs and new virus-based materials for the delivery of drugs or nucleic acid-based therapies. Here we report the assembly of capsid proteins of the cowpea chlorotic mottle virus (CCMV) around DNA into defined structures at neutral pH. Depending on the type of DNA used, we are able to create spherical structures of various diameters and rods of various lengths. In order to determine the shape dependency, the cellular uptake routes and intracellular positioning of these formed polymorphic VLPs in RAW264.7, HeLa and HEK 293 cells are evaluated using flow cytometry analysis with specific chemical inhibitors for different uptake routes. We observed particular uptake routes for the various CCMV-based nanostructures, but the experiments point to clathrin-mediated endocytosis as the major route for cell entry for the studied VLPs. Confocal microscopy reveals that the formed VLPs enter the cells, with clear colocalization in the endosomes. The obtained results provide insight in the cargo dependent VLP morphology and increase the understanding of shape dependent uptake into cells, which is relevant in the design of new virus-based structures with applications in drug and gene delivery.
- Subjects :
- Cytochalasin D
Endosome
Chlorpromazine
viruses
UT-Hybrid-D
Pharmaceutical Science
Plant virus
02 engineering and technology
Gene delivery
Endocytosis
Transfection
Virus
03 medical and health sciences
Mice
Animals
Humans
030304 developmental biology
Cowpea chlorotic mottle virus
0303 health sciences
biology
Chemistry
HEK 293 cells
DNA
Virus-like particles
021001 nanoscience & nanotechnology
biology.organism_classification
Bromovirus
3. Good health
Nanostructures
HEK293 Cells
RAW 264.7 Cells
Capsid
Cargo controlled assembly
Biophysics
Capsid Proteins
0210 nano-technology
HeLa Cells
Subjects
Details
- Language :
- English
- ISSN :
- 01683659
- Database :
- OpenAIRE
- Journal :
- Journal of controlled release, 307, 342-354. Elsevier
- Accession number :
- edsair.doi.dedup.....1cea6d98edd6e127c61cb07de59a581b