Massively Parallelized Molecular Force Manipulation with On-Demand Thermal and Optical Control

Su, Hanquan; Brockman, Joshua M.; Duan, Yuxin; Sen, Navoneel; Chhabra, Hemani; Bazrafshan, Alisina; Blanchard, Aaron T.; Meyer, Travis A.; Andrews, Brooke A; Doye, Jonathan P. K.; Ke, Yonggang; Dyer, R. Brian; Salaita, Khalid

doi:10.1021/jacs.1c08796

Cited by 7 publications

(8 citation statements)

References 53 publications

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“…The treated cells showed ∼50% less spread area and ∼80% less tension signal compared to untreated cells. This confirms that the reported tension is reversible and mediated by F-actin and myosin activity in alignment with prior literature. − Note that the rate of hairpin closing ( k close ) is exponentially dependent on the magnitude of applied force, and as the external force drops a few pN below F 1/2 , refolding rates will approach the ms time scale . Thus, the tension signal decreased immediately upon inhibitor treatment.…”

Section: Resultssupporting

confidence: 89%

“…60−62 Note that the rate of hairpin closing (k close ) is exponentially dependent on the magnitude of applied force, and as the external force drops a few pN below F 1/2 , refolding rates will approach the ms time scale. 63 Thus, the tension signal decreased immediately upon inhibitor treatment. HeLa Cell Integrins Generate F < 19 pN on 13 kPa Hydrogels.…”

Section: ■ Results and Discussionmentioning

confidence: 94%

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All-Covalent Nuclease-Resistant and Hydrogel-Tethered DNA Hairpin Probes Map pN Cell Traction Forces

Rashid,

Dong,

Ogasawara

et al. 2023

ACS Appl. Mater. Interfaces

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Cells sense and respond to the physical properties of their environment through receptor-mediated signaling, a process known as mechanotransduction, which can modulate critical cellular functions such as proliferation, differentiation, and survival. At the molecular level, cell adhesion receptors, such as integrins, transmit piconewton (pN)-scale forces to the extracellular matrix, and the magnitude of the force plays a critical role in cell signaling. The most sensitive approach to measuring integrin forces involves DNA hairpin-based sensors, which are used to quantify and map forces in living cells. Despite the broad use of DNA hairpin sensors to study a variety of mechanotransduction processes, these sensors are typically anchored to rigid glass slides, which are orders of magnitude stiffer than the extracellular matrix and hence modulate native biological responses. Here, we have developed nuclease-resistant DNA hairpin probes that are all covalently tethered to PEG hydrogels to image cell traction forces on physiologically relevant substrate stiffness. Using HeLa cells as a model cell line, we show that the molecular forces transmitted by integrins are highly sensitive to the bulk modulus of the substrate, and cells cultured on the 6 and 13 kPa gels produced a greater number of hairpin unfolding events compared to the 2 kPa substrates. Tension signals are spatially colocalized with pY118-paxillin, confirming focal adhesion-mediated probe opening. Additionally, we found that integrin forces are greater than 5.8 pN but less than 19 pN on 13 kPa gels. This work provides a general strategy to integrate molecular tension probes into hydrogels, which can better mimic in vivo mechanotransduction.

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Section: Resultssupporting

confidence: 89%

Section: ■ Results and Discussionmentioning

confidence: 94%

All-Covalent Nuclease-Resistant and Hydrogel-Tethered DNA Hairpin Probes Map pN Cell Traction Forces

Rashid,

Dong,

Ogasawara

et al. 2023

ACS Appl. Mater. Interfaces

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“…In particular, we used the second version of the model that has been fine-tuned to improve the modelling of origami structure [27]. As a results of its accurate description of the structural [28] and mechanical [29] properties of DNA origami, it has been widely used in the field of DNA nanotechnology, particularly to describe origamis with increasingly functionally complex properties that are otherwise hard to predict [11,[30][31][32][33][34][35][36]. The oxDNA model is particularly well-suited to study jointed origami, as it can also capture the important local features of the junction, e.g., an accurate description of the mechanical properties of single-stranded DNA [26] is important to model the role of the single-stranded linkers on joint flexibility.…”

Section: Methodsmentioning

confidence: 99%

“…One area of increasing interest is their use as nanoscale mechanical devices [7]. In this field of DNA "mechanotechnology" [8] DNA constructs are used to generate [9][10][11], transmit [12] and sense [13] nanoscale forces. One approach is through the use of flexible DNA origami that can undergo large-scale structural changes [14][15][16]; for example, an externally-actuated shape change could be used to apply a force [17], or a force could be sensed through the change in structure it induces [18][19][20].…”

Section: Introductionmentioning

confidence: 99%

The Free-Energy Landscape of a Mechanically Bistable DNA Origami

Wong

Doye

2022

Applied Sciences

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Molecular simulations using coarse-grained models allow the structure, dynamics and mechanics of DNA origamis to be comprehensively characterized. Here, we focus on the free-energy landscape of a jointed DNA origami that has been designed to exhibit two mechanically stable states and for which a bistable landscape has been inferred from ensembles of structures visualized by electron microscopy. Surprisingly, simulations using the oxDNA model predict that the defect-free origami has a single free-energy minimum. The expected second state is not stable because the hinge joints do not simply allow free angular motion but instead lead to increasing free-energetic penalties as the joint angles relevant to the second state are approached. This raises interesting questions about the cause of this difference between simulations and experiment, such as how assembly defects might affect the ensemble of structures observed experimentally.

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“…Tools for manipulating signaling, activity, and function of specific cells in living organisms have enormous potential to promote novel perturbation biology approaches and unprecedented therapeutic strategies. − The past decades have witnessed a drastic expansion in physical perturbation methods including nano-/microelectrode arrays, , microfluidics, , optogenetics, − upconversion nanoparticles, − thermogenetics, − sonogenetics, , and mechanogenetics, , enabling neuromodulation, stem cell differentiation, immune cell activation, and cell migration and adhesion with high spatiotemporal precision. However, despite their potential, broad in vivo applications of these tools have been lagging because of technical limitations.…”

mentioning

confidence: 99%

Hydrogel Magnetomechanical Actuator Nanoparticles for Wireless Remote Control of Mechanosignaling In Vivo

et al. 2023

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As a new enabling nanotechnology tool for wireless, target-specific, and long-distance stimulation of mechanoreceptors in vivo, here we present a hydrogel magnetomechanical actuator (h-MMA) nanoparticle. To allow both deep-tissue penetration of input signals and efficient force generation, h-MMA integrates a two-step transduction mechanism that converts magnetic anisotropic energy to thermal energy within its magnetic core (i.e., Zn0.4Fe2.6O4 nanoparticle cluster) and then to mechanical energy to induce the surrounding polymer (i.e., pNiPMAm) shell contraction, finally delivering forces to activate targeted mechanoreceptors. We show that h-MMAs enable on-demand modulation of Notch signaling in both fluorescence reporter cell lines and a xenograft mouse model, demonstrating its utility as a powerful in vivo perturbation approach for mechanobiology interrogation in a minimally invasive and untethered manner.

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Massively Parallelized Molecular Force Manipulation with On-Demand Thermal and Optical Control

Cited by 7 publications

References 53 publications

All-Covalent Nuclease-Resistant and Hydrogel-Tethered DNA Hairpin Probes Map pN Cell Traction Forces

All-Covalent Nuclease-Resistant and Hydrogel-Tethered DNA Hairpin Probes Map pN Cell Traction Forces

The Free-Energy Landscape of a Mechanically Bistable DNA Origami

Hydrogel Magnetomechanical Actuator Nanoparticles for Wireless Remote Control of Mechanosignaling In Vivo

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