IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v8y2017i1d10.1038_ncomms15334.html
   My bibliography  Save this article

Hit and go CAS9 delivered through a lentiviral based self-limiting circuit

Author

Listed:
  • Gianluca Petris

    (Centre for Integrative Biology (CIBIO), University of Trento, Laboratory of Molecular Virology)

  • Antonio Casini

    (Centre for Integrative Biology (CIBIO), University of Trento, Laboratory of Molecular Virology)

  • Claudia Montagna

    (Centre for Integrative Biology (CIBIO), University of Trento, Laboratory of Molecular Virology)

  • Francesca Lorenzin

    (Centre for Integrative Biology (CIBIO), University of Trento, Laboratory of Computational Oncology)

  • Davide Prandi

    (Centre for Integrative Biology (CIBIO), University of Trento, Laboratory of Computational Oncology)

  • Alessandro Romanel

    (Centre for Integrative Biology (CIBIO), University of Trento, Laboratory of Computational Oncology)

  • Jacopo Zasso

    (Centre for Integrative Biology (CIBIO), Laboratory of Stem Cell Biology)

  • Luciano Conti

    (Centre for Integrative Biology (CIBIO), Laboratory of Stem Cell Biology)

  • Francesca Demichelis

    (Centre for Integrative Biology (CIBIO), University of Trento, Laboratory of Computational Oncology)

  • Anna Cereseto

    (Centre for Integrative Biology (CIBIO), University of Trento, Laboratory of Molecular Virology)

Abstract

In vivo application of the CRISPR-Cas9 technology is still limited by unwanted Cas9 genomic cleavages. Long-term expression of Cas9 increases the number of genomic loci non-specifically cleaved by the nuclease. Here we develop a Self-Limiting Cas9 circuit for Enhanced Safety and specificity (SLiCES) which consists of an expression unit for Streptococcus pyogenes Cas9 (SpCas9), a self-targeting sgRNA and a second sgRNA targeting a chosen genomic locus. The self-limiting circuit results in increased genome editing specificity by controlling Cas9 levels. For its in vivo utilization, we next integrate SLiCES into a lentiviral delivery system (lentiSLiCES) via circuit inhibition to achieve viral particle production. Upon delivery into target cells, the lentiSLiCES circuit switches on to edit the intended genomic locus while simultaneously stepping up its own neutralization through SpCas9 inactivation. By preserving target cells from residual nuclease activity, our hit and go system increases safety margins for genome editing.

Suggested Citation

  • Gianluca Petris & Antonio Casini & Claudia Montagna & Francesca Lorenzin & Davide Prandi & Alessandro Romanel & Jacopo Zasso & Luciano Conti & Francesca Demichelis & Anna Cereseto, 2017. "Hit and go CAS9 delivered through a lentiviral based self-limiting circuit," Nature Communications, Nature, vol. 8(1), pages 1-9, August.
  • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms15334
    DOI: 10.1038/ncomms15334
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/ncomms15334
    File Function: Abstract
    Download Restriction: no

    File URL: https://libkey.io/10.1038/ncomms15334?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
    ---><---

    Citations

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


    Cited by:

    1. Raed Ibraheim & Phillip W. L. Tai & Aamir Mir & Nida Javeed & Jiaming Wang & Tomás C. Rodríguez & Suk Namkung & Samantha Nelson & Eraj Shafiq Khokhar & Esther Mintzer & Stacy Maitland & Zexiang Chen &, 2021. "Self-inactivating, all-in-one AAV vectors for precision Cas9 genome editing via homology-directed repair in vivo," Nature Communications, Nature, vol. 12(1), pages 1-17, 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:8:y:2017:i:1:d:10.1038_ncomms15334. 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.