IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v13y2022i1d10.1038_s41467-022-34269-7.html
   My bibliography  Save this article

High-affinity chromodomains engineered for improved detection of histone methylation and enhanced CRISPR-based gene repression

Author

Listed:
  • G. Veggiani

    (The Anvil Institute
    Louisiana State University)

  • R. Villaseñor

    (Ludwig-Maximilians-University
    University of Zurich)

  • G. D. Martyn

    (The Anvil Institute
    University of Waterloo)

  • J. Q. Tang

    (The Anvil Institute
    University of Waterloo)

  • M. W. Krone

    (University of North Carolina at Chapel Hill, CB 3290)

  • J. Gu

    (The Anvil Institute)

  • C. Chen

    (The Anvil Institute)

  • M. L. Waters

    (University of North Carolina at Chapel Hill, CB 3290)

  • K. H. Pearce

    (University of North Carolina at Chapel Hill)

  • T. Baubec

    (University of Zurich
    Utrecht University)

  • S. S. Sidhu

    (The Anvil Institute
    University of Waterloo)

Abstract

Histone methylation is an important post-translational modification that plays a crucial role in regulating cellular functions, and its dysregulation is implicated in cancer and developmental defects. Therefore, systematic characterization of histone methylation is necessary to elucidate complex biological processes, identify biomarkers, and ultimately, enable drug discovery. Studying histone methylation relies on the use of antibodies, but these suffer from lot-to-lot variation, are costly, and cannot be used in live cells. Chromatin-modification reader domains are potential affinity reagents for methylated histones, but their application is limited by their modest affinities. We used phage display to identify key residues that greatly enhance the affinities of Cbx chromodomains for methylated histone marks and develop a general strategy for enhancing the affinity of chromodomains of the human Cbx protein family. Our strategy allows us to develop powerful probes for genome-wide binding analysis and live-cell imaging. Furthermore, we use optimized chromodomains to develop extremely potent CRISPR-based repressors for tailored gene silencing. Our results highlight the power of engineered chromodomains for analyzing protein interaction networks involving chromatin and represent a modular platform for efficient gene silencing.

Suggested Citation

  • G. Veggiani & R. Villaseñor & G. D. Martyn & J. Q. Tang & M. W. Krone & J. Gu & C. Chen & M. L. Waters & K. H. Pearce & T. Baubec & S. S. Sidhu, 2022. "High-affinity chromodomains engineered for improved detection of histone methylation and enhanced CRISPR-based gene repression," Nature Communications, Nature, vol. 13(1), pages 1-15, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-34269-7
    DOI: 10.1038/s41467-022-34269-7
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/s41467-022-34269-7
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/s41467-022-34269-7?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. Cristiana Lungu & Sabine Pinter & Julian Broche & Philipp Rathert & Albert Jeltsch, 2017. "Modular fluorescence complementation sensors for live cell detection of epigenetic signals at endogenous genomic sites," Nature Communications, Nature, vol. 8(1), pages 1-13, December.
    2. Mike V. Van & Taihei Fujimori & Lacramioara Bintu, 2021. "Nanobody-mediated control of gene expression and epigenetic memory," Nature Communications, Nature, vol. 12(1), pages 1-12, December.
    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.

      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:13:y:2022:i:1:d:10.1038_s41467-022-34269-7. 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.