IDEAS home Printed from https://ideas.repec.org/a/nat/nature/v519y2015i7542d10.1038_nature14245.html
   My bibliography  Save this article

Cas9 specifies functional viral targets during CRISPR–Cas adaptation

Author

Listed:
  • Robert Heler

    (Laboratory of Bacteriology, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA)

  • Poulami Samai

    (Laboratory of Bacteriology, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA)

  • Joshua W. Modell

    (Laboratory of Bacteriology, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA)

  • Catherine Weiner

    (Laboratory of Bacteriology, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA)

  • Gregory W. Goldberg

    (Laboratory of Bacteriology, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA)

  • David Bikard

    (Laboratory of Bacteriology, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA
    Synthetic Biology Group, Institut Pasteur, 28 Rue du Dr. Roux, 75015 Paris, France)

  • Luciano A. Marraffini

    (Laboratory of Bacteriology, The Rockefeller University, 1230 York Avenue, New York, New York 10065, USA)

Abstract

Clustered regularly interspaced short palindromic repeat (CRISPR) loci and their associated (Cas) proteins provide adaptive immunity against viral infection in prokaryotes. Upon infection, short phage sequences known as spacers integrate between CRISPR repeats and are transcribed into small RNA molecules that guide the Cas9 nuclease to the viral targets (protospacers). Streptococcus pyogenes Cas9 cleavage of the viral genome requires the presence of a 5′-NGG-3′ protospacer adjacent motif (PAM) sequence immediately downstream of the viral target. It is not known whether and how viral sequences flanked by the correct PAM are chosen as new spacers. Here we show that Cas9 selects functional spacers by recognizing their PAM during spacer acquisition. The replacement of cas9 with alleles that lack the PAM recognition motif or recognize an NGGNG PAM eliminated or changed PAM specificity during spacer acquisition, respectively. Cas9 associates with other proteins of the acquisition machinery (Cas1, Cas2 and Csn2), presumably to provide PAM-specificity to this process. These results establish a new function for Cas9 in the genesis of prokaryotic immunological memory.

Suggested Citation

  • Robert Heler & Poulami Samai & Joshua W. Modell & Catherine Weiner & Gregory W. Goldberg & David Bikard & Luciano A. Marraffini, 2015. "Cas9 specifies functional viral targets during CRISPR–Cas adaptation," Nature, Nature, vol. 519(7542), pages 199-202, March.
    • Handle: RePEc:nat:nature:v:519:y:2015:i:7542:d:10.1038_nature14245
    DOI: 10.1038/nature14245
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/nature14245
    File Function: Abstract
    Download Restriction: Access to the full text of the articles in this series is restricted.
    File URL: https://libkey.io/10.1038/nature14245?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
    ---><---

    As the access to this document is restricted, you may want to search for a different version of it.

    Citations

    Citations are extracted by the CitEc Project, subscribe to its RSS feed for this item.
    as


    Cited by:

    1. Grace N. Hibshman & Jack P. K. Bravo & Matthew M. Hooper & Tyler L. Dangerfield & Hongshan Zhang & Ilya J. Finkelstein & Kenneth A. Johnson & David W. Taylor, 2024. "Unraveling the mechanisms of PAMless DNA interrogation by SpRY-Cas9," 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:nature:v:519:y:2015:i:7542:d:10.1038_nature14245. 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.

    We have no bibliographic references for this item. You can help adding them by using 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.