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Programmable Multivalent DNA-Origami Tension Probes for Reporting Cellular Traction Forces
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posted on 2018-06-18, 00:00 authored by Palash
K. Dutta, Yun Zhang, Aaron T. Blanchard, Chenghao Ge, Muaz Rushdi, Kristin Weiss, Cheng Zhu, Yonggang Ke, Khalid SalaitaMechanical
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.
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tailorable numberDNA origami tension sensorReporting Cellular Traction Forces Mechanical forcesprotein assembliesDOTP-functionalized surfaces increasesProgrammable Multivalent DNA-Origami Tension ProbesDOTP librarysingle-molecule force spectroscopyblood plateletssix-helix-bundle DNA-origami tension probestension signalnanoscale organizationtension-reporting hairpinsnanometer-length scaletunable tension response thresholdtunable numbernovel libraryforce-sensing probesforce generationcell-receptor ligandsligand densitycell development
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