IDEAS home Printed from https://ideas.repec.org/a/nat/natcom/v8y2017i1d10.1038_ncomms15790.html
   My bibliography  Save this article

Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery

Author

Listed:
  • Holly A. Rees

    (Harvard University
    Howard Hughes Medical Institute, Harvard University
    Broad Institute of MIT and Harvard)

  • Alexis C. Komor

    (Harvard University
    Howard Hughes Medical Institute, Harvard University
    Broad Institute of MIT and Harvard)

  • Wei-Hsi Yeh

    (Harvard University
    Howard Hughes Medical Institute, Harvard University
    Broad Institute of MIT and Harvard
    Eaton-Peabody Laboratory, Massachusetts Eye and Ear Infirmary)

  • Joana Caetano-Lopes

    (Orthopaedic Research Laboratories, Boston Children’s Hospital
    Harvard Medical School)

  • Matthew Warman

    (Orthopaedic Research Laboratories, Boston Children’s Hospital
    Harvard Medical School)

  • Albert S. B. Edge

    (Eaton-Peabody Laboratory, Massachusetts Eye and Ear Infirmary
    Program in Speech and Hearing Bioscience and Technology, Harvard Medical School
    Harvard Medical School)

  • David R. Liu

    (Harvard University
    Howard Hughes Medical Institute, Harvard University
    Broad Institute of MIT and Harvard)

Abstract

We recently developed base editing, a genome-editing approach that enables the programmable conversion of one base pair into another without double-stranded DNA cleavage, excess stochastic insertions and deletions, or dependence on homology-directed repair. The application of base editing is limited by off-target activity and reliance on intracellular DNA delivery. Here we describe two advances that address these limitations. First, we greatly reduce off-target base editing by installing mutations into our third-generation base editor (BE3) to generate a high-fidelity base editor (HF-BE3). Next, we purify and deliver BE3 and HF-BE3 as ribonucleoprotein (RNP) complexes into mammalian cells, establishing DNA-free base editing. RNP delivery of BE3 confers higher specificity even than plasmid transfection of HF-BE3, while maintaining comparable on-target editing levels. Finally, we apply these advances to deliver BE3 RNPs into both zebrafish embryos and the inner ear of live mice to achieve specific, DNA-free base editing in vivo.

Suggested Citation

  • Holly A. Rees & Alexis C. Komor & Wei-Hsi Yeh & Joana Caetano-Lopes & Matthew Warman & Albert S. B. Edge & David R. Liu, 2017. "Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery," Nature Communications, Nature, vol. 8(1), pages 1-10, August.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_ncomms15790
    DOI: 10.1038/ncomms15790
    as

    Download full text from publisher

    File URL: https://www.nature.com/articles/ncomms15790
    File Function: Abstract
    Download Restriction: no
    File URL: https://libkey.io/10.1038/ncomms15790?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. Catarina Rebelo & Tiago Reis & Joana Guedes & Cláudia Saraiva & Artur Filipe Rodrigues & Susana Simões & Liliana Bernardino & João Peça & Sónia L. C. Pinho & Lino Ferreira, 2022. "Efficient spatially targeted gene editing using a near-infrared activatable protein-conjugated nanoparticle for brain applications," Nature Communications, Nature, vol. 13(1), pages 1-16, December.
    2. Jing Guo & Luyao Gong & Haiying Yu & Ming Li & Qiaohui An & Zhenquan Liu & Shuru Fan & Changjialian Yang & Dahe Zhao & Jing Han & Hua Xiang, 2024. "Engineered minimal type I CRISPR-Cas system for transcriptional activation and base editing in human cells," Nature Communications, Nature, vol. 15(1), pages 1-16, 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_ncomms15790. 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.