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  • Correspondence
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Failure to detect DNA-guided genome editing using Natronobacterium gregoryi Argonaute

Nature Biotechnology volume 35, pages 17–18 (2017)Cite this article

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To the Editor:

DNA-guided DNA cleavage using Argonaute-family proteins was previously reported but of limited practical utility due to the requirement for supraphysiological conditions, such as high temperature1,2. Recently, Gao et al.3 reported an Argonaute protein isolated from Natronobacterium gregoryi (NgAgo) as a genome engineering tool for editing the human genome. After transfecting human cell lines with a plasmid DNA encoding NgAgo and a 24-nucleotide, 5'-phosphorylated single-strand guide DNA (gDNA), they showed gene editing at endogenous targets. Here, we report the results of three different groups (Cathomen, Supplementary Fig. 1 and Supplementary Methods 1; Ekker, Supplementary Fig. 2 and Supplementary Methods 2; and Kim, Supplementary Fig. 3 and Supplementary Methods 3), independently attempting to reproduce the original findings of Gao et al.3, specifically focusing on the evidence for DNA edits in cultured human cell lines. All three groups synthesized the same 5'-phosphorylated gDNA sequences and used the NgAgo vector provided by Gao et al.3 via Addgene (Cambridge, MA, USA) to transfect the same cell lines, and analyzed the genomic DNA for signs of gene editing. Controls confirmed efficient delivery of both plasmid DNA and gDNAs as well as expression of the NgAgo protein. Despite various attempts to optimize NgAgo-mediated genome editing in three of the reported cell lines, no evidence of successful editing of endogenous target sequences was detected.

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Change history

  • 02 August 2017

    The Article 'DNA-guided genome editing using the Natronobacterium gregoryi Argonaute' by F. Gao et al. (Nature Biotechnology, 34, 768-773, 2016) on which this Correspondence is based was retracted by the authors on 2 August 2017.

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Acknowledgements

We thank all members of our laboratories who made valuable contributions in trying to establish NgAgo-based genome editing. This work was supported by grants from the German Federal Ministry of Education and Research (BMBF-01EO0803 to T.C.), American Heart Association (16PRE30470004 to H.A.), the National Institutes of Health (GM63904 to S.C.E.), the Mayo Foundation (to H.A., S.N., Z.L., and S.C.E.), and the Institute for Basic Science (IBS-R021-D1 to J.-S.K.).

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Author notes

  1. Seung Hwan Lee, Giandomenico Turchiano and Hirotaka Ata: These authors contributed equally to this work.

Authors and Affiliations

  1. Center for Genome Engineering, Institute for Basic Science (IBS), Seoul, South Korea

    Seung Hwan Lee, Seuk-Min Ryu & Jin-Soo Kim

  2. Institute for Cell and Gene Therapy, Medical Center—University of Freiburg, Freiburg, Germany

    Giandomenico Turchiano, Marianna Romito & Toni Cathomen

  3. Center for Chronic Immunodeficiency, Medical Center—University of Freiburg, Freiburg, Germany

    Giandomenico Turchiano, Marianna Romito & Toni Cathomen

  4. Mayo Graduate School, Mayo Clinic, Rochester, Minnesota, USA

    Hirotaka Ata & Somaira Nowsheen

  5. Graduate School, Faculty of Biology, University of Freiburg, Freiburg, Germany

    Marianna Romito

  6. Division of Oncology Research, Mayo Clinic, Rochester, Minnesota, USA

    Zhenkun Lou

  7. Department of Chemistry, Seoul National University, Seoul, South Korea

    Seuk-Min Ryu & Jin-Soo Kim

  8. Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota, USA

    Stephen C Ekker

  9. Faculty of Medicine, University of Freiburg, Freiburg, Germany

    Toni Cathomen

Authors
  1. Seung Hwan Lee
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  10. Jin-Soo Kim
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Corresponding authors

Correspondence to Stephen C Ekker, Toni Cathomen or Jin-Soo Kim.

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Competing interests

S.C.E. is a co-founder of Lifengine Technologies, Inc.; T.C. is a consultant for TRACR Hematology; J.-S.K. is a co-founder and shareholder of ToolGen, Inc.

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Editor's note: This article has been peer-reviewed.

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Supplementary Text and Figures

Supplementary Figures 1–3 and Supplementary Materials 1–3 (PDF 4220 kb)

Supplementary Dataset

Supplementary Dataset 1 (ZIP 1839 kb)

Supplementary Dataset

Supplementary Dataset 2 (ZIP 11465 kb)

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Lee, S., Turchiano, G., Ata, H. et al. Failure to detect DNA-guided genome editing using Natronobacterium gregoryi Argonaute. Nat Biotechnol 35, 17–18 (2017). https://doi.org/10.1038/nbt.3753

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