Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Correspondence
  • Published:

To cleave or not to cleave: therapeutic gene editing with and without programmable nucleases

Nature Reviews Drug Discovery volume 16, page 296 (2017)Cite this article

Subjects

Access through your institution
Buy or subscribe

In recent years, pioneering clinical studies involving the use of programmable nucleases to achieve gene editing have begun to evaluate the therapeutic potential of such approaches. For example, Sangamo BioSciences has reported successful proof-of-concept clinical studies to treat HIV infection using an engineered zinc-finger nuclease (ZFN) to inactivate the gene coding for CC chemokine receptor 5 (CCR5) —a co-receptor for HIV — in T cells ex vivo, as highlighted in a news article in this journal1 (Nat. Rev. Drug Discov. 13, 321–322; 2014). Initial clinical trials using CRISPR–Cas9 gene editing technology, which has rapidly become a widely used gene editing platform in biomedical research2, were also announced in 2016.

In this article, however, we focus on an alternative group of platforms for achieving therapeutic gene editing without the use of programmable nucleases. Although such approaches were pioneered in the 1990s3, before the advent of the nuclease-based gene editing platforms, their use and progression to clinical trials since has been held back by issues including frequencies of gene editing that are typically well below those needed for potential therapeutic applications. However, in the past 2 years, results with a diverse group of gene editing platforms — ranging from recombinogenic adeno-associated viral (AAV) vectors, to various oligonucleotide-only approaches, to nucleobase modification (Table 1) — have demonstrated substantially improved gene editing efficiencies4,5,6,7,8,9, spurring renewed interest in such approaches. Moreover, these platforms do not require cleavage of the targeted DNA to achieve gene editing — a characteristic of nuclease-based mechanisms that has raised concerns about random insertion and deletions (indels) that occur in some proportion of the targeted cleavage sites and their potential for off-target genotoxicity. Here, after briefly noting some of the history of the field, we highlight recent advances with several gene editing platforms that do not use nucleases.

Table 1 Major therapeutic genome editing approaches
Full size table

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

References

  1. Owens, B. Zinc-finger nucleases make the cut in HIV. Nat. Rev. Drug Discov. 13, 321–322 (2014).

    Article  Google Scholar 

  2. Cox, D. B., Platt, R. J. & Zhang, F. Therapeutic genome editing: prospects and challenges. Nat. Med. 21, 121–131 (2015).

    Article  CAS  Google Scholar 

  3. Woolf, T. M. Therapeutic repair of mutated nucleic acid sequences. Nat. Biotechnol. 16, 341–344 (1998).

    Article  CAS  Google Scholar 

  4. Barzel, A. et al. Promoterless gene targeting without nucleases ameliorates haemophilia B in mice. Nature 517, 360–364 (2015).

    Article  CAS  Google Scholar 

  5. McNeer, N. A. et al. Nanoparticles that deliver triplex-forming peptide nucleic acid molecules correct F508del CFTR in airway epithelium. Nat. Commun. 6, 6952 (2015).

    Article  CAS  Google Scholar 

  6. Bahal, R. et al. In vivo correction of anaemia in β-thalassemic mice by γPNA-mediated gene editing with nanoparticle delivery. Nat. Commun. 7, 13304 (2016).

    Article  CAS  Google Scholar 

  7. van Ravesteyn, T. W. et al. LNA modification of single-stranded DNA oligonucleotides allows subtle gene modification in mismatch-repair-proficient cells. Proc. Natl Acad. Sci. USA 113, 4122–4127 (2016).

    Article  CAS  Google Scholar 

  8. Komor, A. C., Kim, Y. B., Packer, M. S., Zuris, J. A. & Liu, D. R. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature 533, 420–424 (2016).

    Article  CAS  Google Scholar 

  9. Yang, L. et al. Engineering and optimising deaminase fusions for genome editing. Nat. Commun. 7, 13330 (2016).

    Article  CAS  Google Scholar 

  10. Engstrom, J. U., Suzuki, T. & Kmiec, E. B. Regulation of targeted gene repair by intrinsic cellular processes. Bioessays 31, 159–168 (2009).

    Article  CAS  Google Scholar 

  11. Rios, X. et al. Stable gene targeting in human cells using single-strand oligonucleotides with modified bases. PLoS ONE 7, e36697 (2012).

    Article  CAS  Google Scholar 

  12. Bertoni, C., Rustagi, A. & Rando, T. A. Enhanced gene repair mediated by methyl-CpG-modified single-stranded oligonucleotides. Nucleic Acids Res. 37, 7468–7482 (2009).

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. ETAGEN Pharma, Inc., Cambridge, 01239, Massachusetts, USA

    Tod M. Woolf

  2. Mouse Engineering Core Facility, University of Nebraska Medical Center, Omaha, 68198, Nebraska, USA

    Channabasavaiah B. Gurumurthy

  3. Massachusetts General Hospital, Cambridge, 02139, Massachusetts, USA

    Frederick Boyce

  4. Gene Editing Institute for Translational Cancer Research, Helen F. Graham Cancer Center, and Research Institute, Newark, 19713, Delaware, USA

    Eric B. Kmiec

Authors
  1. Tod M. Woolf
    View author publications

    Search author on:PubMed Google Scholar

  2. Channabasavaiah B. Gurumurthy
    View author publications

    Search author on:PubMed Google Scholar

  3. Frederick Boyce
    View author publications

    Search author on:PubMed Google Scholar

  4. Eric B. Kmiec
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to Tod M. Woolf.

Ethics declarations

Competing interests

T.M.W., F.B. and E.B.K. own shares in ETAGEN Pharma, where T.M.W. served as chief executive officer, and F.B. and E.B.K. serve as scientific advisors. T.M.W. began a position as a technology licensing officer at the Massachusetts Institute of Technology (MIT) Technology Licensing Office on 9 November 2016, after the submission of this article, but before the final post-submission editing was completed. T.M.W. does not work on licensing of CRISPR–Cas9 at MIT and does not have any financial interest in CRISPR–Cas9 at MIT.

Related links

FURTHER INFORMATION

US patent 7,258,854

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Woolf, T., Gurumurthy, C., Boyce, F. et al. To cleave or not to cleave: therapeutic gene editing with and without programmable nucleases. Nat Rev Drug Discov 16, 296 (2017). https://doi.org/10.1038/nrd.2017.42

Download citation

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/nrd.2017.42

Search

Advanced search

Quick links

Nature Briefing: Translational Research

Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology, drug discovery and pharma.

Get what matters in translational research, free to your inbox weekly. Sign up for Nature Briefing: Translational Research