<div class="csl-bib-body">
<div class="csl-entry">Majkova, A., Hellmeier, J., Platzer, R., Huppa, J., & Sevcsik, E. (2023, August 29). <i>Designing functionalized DNA origami-based biointerfaces for probing T-cell activation</i> [Poster Presentation]. 12th Single Molecule Localization Microscopy Symposium (SMLMS) 2023, Wien, Austria. http://hdl.handle.net/20.500.12708/188938</div>
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dc.identifier.uri
http://hdl.handle.net/20.500.12708/188938
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dc.description.abstract
The DNA origami technique [1] allows to create flexible scaffolds for the organization of single biomolecules with nanometer precision, yielding powerful tools to study cellular processes on the molecular level. Here, we apply DNA origami to investigate the earliest molecular events of T-cell activation, a step required to trigger a complex immune response.
We design, produce, and characterize DNA origami nanostructures (namely 70x100 nm 2D platforms) to engineer a biointerface that mimics an antigen-presenting cell (APC). DNA origami platforms are further functionalized with proteins, allowing us to manipulate the organization of T-cell receptor ligands and other proteins at the APC interface. Functionalized DNA origami are attached to supported lipid bilayers (SLBs) via cholesterol strands, so platforms can diffuse freely.
We use single molecule fluorescence microscopy to evaluate platform folding, functionalization with proteins (two-color colocalization), and mobility (single molecule tracking). In the final step, we bring fabricated biointerfaces in contact with primary murine T-cells, to investigate the effect of various platform designs on T-cell interaction and signalling.
To this date, we have evaluated different functionalization strategies for the site-specific decoration of DNA origami platforms with TCR ligands. The strategy employing monovalent streptavidin and a biotinylated ligand yielded high efficiency, high protein functionality, and a defined stoichiometry [2] while also allowing heterofunctionalization with two or more different biomolecules. In another study, we applied our biointerfaces to determine the minimal signalling unit able to promote T-cell activation or stimulation, suggesting transient T-cell: APC binding [3]. Currently, we design, characterize, and employ heterobifunctional DNA origami platforms to determine the effect of co-stimulatory and adhesion proteins as well as weak agonists, co-, and antagonists on T-cell:APC interaction, aiming to engineer a tool applicable for probing a broad range of cell-cell interactions.
en
dc.language.iso
en
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dc.subject
Biophysics
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dc.subject
DNA origami
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dc.subject
T-cells activation
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dc.title
Designing functionalized DNA origami-based biointerfaces for probing T-cell activation
en
dc.type
Presentation
en
dc.type
Vortrag
de
dc.contributor.affiliation
Max-Planck-Institut für Biochemie
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dc.contributor.affiliation
Medical University of Vienna, Austria
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dc.contributor.affiliation
Medical University of Vienna, Austria
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dc.type.category
Poster Presentation
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tuw.researchTopic.id
M2
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tuw.researchTopic.id
M6
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tuw.researchTopic.name
Materials Characterization
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tuw.researchTopic.name
Biological and Bioactive Materials
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tuw.researchTopic.value
30
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tuw.researchTopic.value
70
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tuw.linking
https://smlms.org/
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tuw.publication.orgunit
E134-04 - Forschungsbereich Biophysics
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tuw.author.orcid
0000-0002-7153-9121
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tuw.author.orcid
0000-0003-2634-8198
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tuw.author.orcid
0000-0002-2155-1675
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tuw.event.name
12th Single Molecule Localization Microscopy Symposium (SMLMS) 2023