IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v14y2023i1d10.1038_s41467-023-38157-6.html
   My bibliography  Save this article

RAD-TGTs: high-throughput measurement of cellular mechanotype via rupture and delivery of DNA tension probes

Author

Listed:
  • Matthew R. Pawlak

    (University of Minnesota)

  • Adam T. Smiley

    (University of Minnesota)

  • Maria Paz Ramirez

    (University of Minnesota)

  • Marcus D. Kelly

    (University of Minnesota)

  • Ghaidan A. Shamsan

    (University of Minnesota)

  • Sarah M. Anderson

    (University of Minnesota)

  • Branden A. Smeester

    (University of Minnesota)

  • David A. Largaespada

    (University of Minnesota)

  • David J. Odde

    (University of Minnesota)

  • Wendy R. Gordon

    (University of Minnesota)

Abstract

Mechanical forces drive critical cellular processes that are reflected in mechanical phenotypes, or mechanotypes, of cells and their microenvironment. We present here “Rupture And Deliver” Tension Gauge Tethers (RAD-TGTs) in which flow cytometry is used to record the mechanical history of thousands of cells exerting forces on their surroundings via their propensity to rupture immobilized DNA duplex tension probes. We demonstrate that RAD-TGTs recapitulate prior DNA tension probe studies while also yielding a gain of fluorescence in the force-generating cell that is detectable by flow cytometry. Furthermore, the rupture propensity is altered following disruption of the cytoskeleton using drugs or CRISPR-knockout of mechanosensing proteins. Importantly, RAD-TGTs can differentiate distinct mechanotypes among mixed populations of cells. We also establish oligo rupture and delivery can be measured via DNA sequencing. RAD-TGTs provide a facile and powerful assay to enable high-throughput mechanotype profiling, which could find various applications, for example, in combination with CRISPR screens and -omics analysis.

Suggested Citation

  • Matthew R. Pawlak & Adam T. Smiley & Maria Paz Ramirez & Marcus D. Kelly & Ghaidan A. Shamsan & Sarah M. Anderson & Branden A. Smeester & David A. Largaespada & David J. Odde & Wendy R. Gordon, 2023. "RAD-TGTs: high-throughput measurement of cellular mechanotype via rupture and delivery of DNA tension probes," Nature Communications, Nature, vol. 14(1), pages 1-11, December.
  • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-38157-6
    DOI: 10.1038/s41467-023-38157-6
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-023-38157-6
    File Function: Abstract
    Download Restriction: no

    File URL: https://libkey.io/10.1038/s41467-023-38157-6?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    References listed on IDEAS

    as
    1. Benjamin L. Bangasser & Ghaidan A. Shamsan & Clarence E. Chan & Kwaku N. Opoku & Erkan Tüzel & Benjamin W. Schlichtmann & Jesse A. Kasim & Benjamin J. Fuller & Brannon R. McCullough & Steven S. Rosenf, 2017. "Shifting the optimal stiffness for cell migration," Nature Communications, Nature, vol. 8(1), pages 1-10, August.
    2. Yun Zhang & Chenghao Ge & Cheng Zhu & Khalid Salaita, 2014. "DNA-based digital tension probes reveal integrin forces during early cell adhesion," Nature Communications, Nature, vol. 5(1), pages 1-10, December.
    3. Matthew J. Paszek & Christopher C. DuFort & Olivier Rossier & Russell Bainer & Janna K. Mouw & Kamil Godula & Jason E. Hudak & Jonathon N. Lakins & Amanda C. Wijekoon & Luke Cassereau & Matthew G. Rub, 2014. "The cancer glycocalyx mechanically primes integrin-mediated growth and survival," Nature, Nature, vol. 511(7509), pages 319-325, July.
    4. Carsten Grashoff & Brenton D. Hoffman & Michael D. Brenner & Ruobo Zhou & Maddy Parsons & Michael T. Yang & Mark A. McLean & Stephen G. Sligar & Christopher S. Chen & Taekjip Ha & Martin A. Schwartz, 2010. "Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics," Nature, Nature, vol. 466(7303), pages 263-266, July.
    Full references (including those not matched with items on IDEAS)

    Most related items

    These are the items that most often cite the same works as this one and are cited by the same works as this one.
    1. Serena Petracchini & Daniel Hamaoui & Anne Doye & Atef Asnacios & Florian Fage & Elisa Vitiello & Martial Balland & Sebastien Janel & Frank Lafont & Mukund Gupta & Benoit Ladoux & Jerôme Gilleron & Te, 2022. "Optineurin links Hace1-dependent Rac ubiquitylation to integrin-mediated mechanotransduction to control bacterial invasion and cell division," Nature Communications, Nature, vol. 13(1), pages 1-22, December.
    2. Yuhang Zhang & Jingyi Du & Xian Liu & Fei Shang & Yunxin Deng & Jiaqing Ye & Yukai Wang & Jie Yan & Hu Chen & Miao Yu & Shimin Le, 2024. "Multi-domain interaction mediated strength-building in human α-actinin dimers unveiled by direct single-molecule quantification," Nature Communications, Nature, vol. 15(1), pages 1-17, December.
    3. Jiankai Wei & Wei Zhang & An Jiang & Hongzhe Peng & Quanyong Zhang & Yuting Li & Jianqing Bi & Linting Wang & Penghui Liu & Jing Wang & Yonghang Ge & Liya Zhang & Haiyan Yu & Lei Li & Shi Wang & Liang, 2024. "Temporospatial hierarchy and allele-specific expression of zygotic genome activation revealed by distant interspecific urochordate hybrids," Nature Communications, Nature, vol. 15(1), pages 1-11, December.
    4. Jérôme R D Soiné & Christoph A Brand & Jonathan Stricker & Patrick W Oakes & Margaret L Gardel & Ulrich S Schwarz, 2015. "Model-based Traction Force Microscopy Reveals Differential Tension in Cellular Actin Bundles," PLOS Computational Biology, Public Library of Science, vol. 11(3), pages 1-16, March.
    5. Mitchell S. Wang & Yuesong Hu & Elisa E. Sanchez & Xihe Xie & Nathan H. Roy & Miguel Jesus & Benjamin Y. Winer & Elizabeth A. Zale & Weiyang Jin & Chirag Sachar & Joanne H. Lee & Yeonsun Hong & Minsoo, 2022. "Mechanically active integrins target lytic secretion at the immune synapse to facilitate cellular cytotoxicity," Nature Communications, Nature, vol. 13(1), pages 1-15, December.
    6. Rahmetullah Varol & Zeynep Karavelioglu & Sevde Omeroglu & Gizem Aydemir & Aslihan Karadag & Hanife E. Meco & Ali A. Demircali & Abdurrahim Yilmaz & Gizem C. Kocal & Gulsum Gencoglan & Muhammed E. Oru, 2022. "Acousto-holographic reconstruction of whole-cell stiffness maps," Nature Communications, Nature, vol. 13(1), pages 1-10, December.
    7. A. Mills & N. Aissaoui & D. Maurel & J. Elezgaray & F. Morvan & J. J. Vasseur & E. Margeat & R. B. Quast & J. Lai Kee-Him & N. Saint & C. Benistant & A. Nord & F. Pedaci & G. Bellot, 2022. "A modular spring-loaded actuator for mechanical activation of membrane proteins," Nature Communications, Nature, vol. 13(1), pages 1-10, December.
    8. Avery Parr & Nicholas R Anderson & Daniel A Hammer, 2019. "A simulation of the random and directed motion of dendritic cells in chemokine fields," PLOS Computational Biology, Public Library of Science, vol. 15(10), pages 1-16, October.
    9. Ehsan Akbari & Melika Shahhosseini & Ariel Robbins & Michael G. Poirier & Jonathan W. Song & Carlos E. Castro, 2022. "Low cost and massively parallel force spectroscopy with fluid loading on a chip," Nature Communications, Nature, vol. 13(1), pages 1-11, December.
    10. Venkat R. Chirasani & Mohammad Ashhar I. Khan & Juilee N. Malavade & Nikolay V. Dokholyan & Brenton D. Hoffman & Sharon L. Campbell, 2023. "Molecular basis and cellular functions of vinculin-actin directional catch bonding," Nature Communications, Nature, vol. 14(1), pages 1-20, December.
    11. Arventh Velusamy & Radhika Sharma & Sk Aysha Rashid & Hiroaki Ogasawara & Khalid Salaita, 2024. "DNA mechanocapsules for programmable piconewton responsive drug delivery," Nature Communications, Nature, vol. 15(1), pages 1-13, December.
    12. Brooke E. Danielsson & Bobin George Abraham & Elina Mäntylä & Jolene I. Cabe & Carl R. Mayer & Anna Rekonen & Frans Ek & Daniel E. Conway & Teemu O. Ihalainen, 2023. "Nuclear lamina strain states revealed by intermolecular force biosensor," Nature Communications, Nature, vol. 14(1), pages 1-15, December.
    13. Myung Hyun Jo & Jing Li & Valentin Jaumouillé & Yuxin Hao & Jessica Coppola & Jiabin Yan & Clare M. Waterman & Timothy A. Springer & Taekjip Ha, 2022. "Single-molecule characterization of subtype-specific β1 integrin mechanics," Nature Communications, Nature, vol. 13(1), pages 1-18, December.
    14. Chrystian Junqueira Alves & Rafael Dariolli & Jonathan Haydak & Sangjo Kang & Theodore Hannah & Robert J. Wiener & Stefanie DeFronzo & Rut Tejero & Gabriele L. Gusella & Aarthi Ramakrishnan & Rodrigo , 2021. "Plexin-B2 orchestrates collective stem cell dynamics via actomyosin contractility, cytoskeletal tension and adhesion," Nature Communications, Nature, vol. 12(1), pages 1-23, December.
    15. Chih-Hao Lu & Kayvon Pedram & Ching-Ting Tsai & Taylor Jones & Xiao Li & Melissa L. Nakamoto & Carolyn R. Bertozzi & Bianxiao Cui, 2022. "Membrane curvature regulates the spatial distribution of bulky glycoproteins," Nature Communications, Nature, vol. 13(1), pages 1-16, December.
    16. Stacy A. Malaker & Nicholas M. Riley & D. Judy Shon & Kayvon Pedram & Venkatesh Krishnan & Oliver Dorigo & Carolyn R. Bertozzi, 2022. "Revealing the human mucinome," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    17. Tianchi Chen & Cecilia H. Fernández-Espartero & Abigail Illand & Ching-Ting Tsai & Yang Yang & Benjamin Klapholz & Pierre Jouchet & Mélanie Fabre & Olivier Rossier & Bianxiao Cui & Sandrine Lévêque-Fo, 2024. "Actin-driven nanotopography promotes stable integrin adhesion formation in developing tissue," Nature Communications, Nature, vol. 15(1), pages 1-16, December.
    18. Sorosh Amiri & Camelia Muresan & Xingbo Shang & Clotilde Huet-Calderwood & Martin A. Schwartz & David A. Calderwood & Michael Murrell, 2023. "Intracellular tension sensor reveals mechanical anisotropy of the actin cytoskeleton," Nature Communications, Nature, vol. 14(1), pages 1-15, December.
    19. Florian Franz & Rafael Tapia-Rojo & Sabina Winograd-Katz & Rajaa Boujemaa-Paterski & Wenhong Li & Tamar Unger & Shira Albeck & Camilo Aponte-Santamaria & Sergi Garcia-Manyes & Ohad Medalia & Benjamin , 2023. "Allosteric activation of vinculin by talin," Nature Communications, Nature, vol. 14(1), pages 1-16, December.
    20. Kaitao Li & Paul Cardenas-Lizana & Jintian Lyu & Anna V. Kellner & Menglan Li & Peiwen Cong & Valencia E. Watson & Zhou Yuan & Eunseon Ahn & Larissa Doudy & Zhenhai Li & Khalid Salaita & Rafi Ahmed & , 2024. "Mechanical force regulates ligand binding and function of PD-1," Nature Communications, Nature, vol. 15(1), pages 1-15, December.

    More about this item

    Statistics

    Access and download statistics

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-38157-6. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    If CitEc recognized a bibliographic reference but did not link an item in RePEc to it, you can help with this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Sonal Shukla or Springer Nature Abstracting and Indexing (email available below). General contact details of provider: http://www.nature.com .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.