10.1021/acs.nanolett.8b01374.s003
Palash
K. Dutta
Palash
K.
Dutta
Yun Zhang
Yun
Zhang
Aaron T. Blanchard
Aaron T.
Blanchard
Chenghao Ge
Chenghao
Ge
Muaz Rushdi
Muaz
Rushdi
Kristin Weiss
Kristin
Weiss
Cheng Zhu
Cheng
Zhu
Yonggang Ke
Yonggang
Ke
Khalid Salaita
Khalid
Salaita
Programmable Multivalent DNA-Origami Tension Probes
for Reporting Cellular Traction Forces
American Chemical Society
2018
tailorable number
DNA origami tension sensor
Reporting Cellular Traction Forces Mechanical forces
protein assemblies
DOTP-functionalized surfaces increases
Programmable Multivalent DNA-Origami Tension Probes
DOTP library
single-molecule force spectroscopy
blood platelets
six-helix-bundle DNA-origami tension probes
tension signal
nanoscale organization
tension-reporting hairpins
nanometer-length scale
tunable tension response threshold
tunable number
novel library
force-sensing probes
force generation
cell-receptor ligands
ligand density
cell development
2018-06-18 00:00:00
Media
https://acs.figshare.com/articles/media/Programmable_Multivalent_DNA-Origami_Tension_Probes_for_Reporting_Cellular_Traction_Forces/6743441
Mechanical
forces are central to most, if not all, biological processes,
including cell development, immune recognition, and metastasis. Because
the cellular machinery mediating mechano-sensing and force generation
is dependent on the nanoscale organization and geometry of protein
assemblies, a current need in the field is the development of force-sensing
probes that can be customized at the nanometer-length scale. In this
work, we describe a DNA origami tension sensor that maps the piconewton
(pN) forces generated by living cells. As a proof-of-concept, we engineered
a novel library of six-helix-bundle DNA-origami tension probes (DOTPs)
with a tailorable number of tension-reporting hairpins (each with
their own tunable tension response threshold) and a tunable number
of cell-receptor ligands. We used single-molecule force spectroscopy
to determine the probes’ tension response thresholds and used
computational modeling to show that hairpin unfolding is semi-cooperative
and orientation-dependent. Finally, we use our DOTP library to map
the forces applied by human blood platelets during initial adhesion
and activation. We find that the total tension signal exhibited by
platelets on DOTP-functionalized surfaces increases with the number
of ligands per DOTP, likely due to increased total ligand density,
and decreases exponentially with the DOTP’s force-response
threshold. This work opens the door to applications for understanding
and regulating biophysical processes involving cooperativity and multivalency.