Abstract

Inefficient gene transfer and low virion concentrations are common limitations of retroviral transduction1. We and others have previously shown that peptides derived from human semen form amyloid fibrils that boost retroviral gene delivery by promoting virion attachment to the target cells2,3,4,5,6,7,8. However, application of these natural fibril-forming peptides is limited by moderate efficiencies, the high costs of peptide synthesis, and variability in fibril size and formation kinetics. Here, we report the development of nanofibrils that self-assemble in aqueous solution from a 12-residue peptide, termed enhancing factor C (EF-C). These artificial nanofibrils enhance retroviral gene transfer substantially more efficiently than semen-derived fibrils or other transduction enhancers. Moreover, EF-C nanofibrils allow the concentration of retroviral vectors by conventional low-speed centrifugation, and are safe and effective, as assessed in an ex vivo gene transfer study. Our results show that EF-C fibrils comprise a highly versatile, convenient and broadly applicable nanomaterial that holds the potential to significantly facilitate retroviral gene transfer in basic research and clinical applications.

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Acknowledgements

The authors thank S. Liu for support in the initial phase of the project and D. Krnavek and M. Schlaier for expert technical assistance. The authors also thank W. Mothes for providing the MLV GAG-CFP plasmid, B. Böhm for Bon cells, J. von Einem for HFF cells, the AIDS Research and Reference Program for U87-MG and TZM-bl cells, and N. Landau for CEMx-M7 (CEMx174 5.25 M7) cells. This work was supported by a grant from the German Research Foundation to J.M. Molecular simulations were performed using the resources of Moscow University Supercomputing Center.

Author information

Author notes

    • Maral Yolamanova
    •  & Christoph Meier

    These authors contributed equally to this work

Affiliations

  1. Institute of Molecular Virology, Ulm University Medical Center, Meyerhofstrasse 1, 89081 Ulm, Germany

    • Maral Yolamanova
    • , Franziska Arnold
    • , Onofrio Zirafi
    • , Shariq M. Usmani
    • , Janis A. Müller
    • , Daniel Sauter
    • , Christine Goffinet
    • , David Palesch
    • , Frank Kirchhoff
    •  & Jan Münch
  2. Institute of Organic Chemistry III/Macromolecular Chemistry, Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany

    • Christoph Meier
    •  & Tanja Weil
  3. Institute of Polymer Science, Ulm University, Albert-Einstein-Allee 47, 89069 Ulm, Germany

    • Alexey K. Shaytan
    • , Pavel G. Khalatur
    •  & Alexei R. Khokhlov
  4. Faculty of Biology, Lomonosov Moscow State University, 1-12 Leninskie Gory, 119991 Moscow, Russia

    • Alexey K. Shaytan
  5. Department of Dermatology and Allergic Diseases, Ulm University Medical Center, Meyerhofstrasse 1, 89081 Ulm, Germany

    • Virag Vas
    •  & Hartmut Geiger
  6. ICREA and Joint BSC-IRB Research Programme in Computational Biology, Institute for Research in Biomedicine, Parc Científic de Barcelona c/ Baldiri Reixac 10, 08028 Barcelona, Spain

    • Carlos W. Bertoncini
    •  & Xavier Salvatella
  7. Central Electron Microscopy Facility, Ulm University, Albert-Einstein-Allee 11, 89069 Ulm, Germany

    • Paul Walther
  8. Department of Urology, Gladstone Institute of Virology and Immunology, University of California at San Francisco, 1650 Owens St, San Francisco, California 94158, USA

    • Nadia R. Roan
  9. Institute of Pharmacology of Natural Products and Clinical Pharmacology, Ulm University, Helmholtzstrasse 20, 89081 Ulm, Germany

    • Oleg Lunov
    •  & Thomas Simmet
  10. Institute for Virology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625 Hannover, Germany

    • Jens Bohne
  11. Institute for Transfusion Medicine, Ulm University and Institute for Clinical Transfusion Medicine and Immunogenetics Ulm, DRK Blood Service Baden-Württemberg – Hessen, gemeinnützige GmbH, Helmholtzstrasse 10, 89081 Ulm, Germany

    • Hubert Schrezenmeier
    •  & Klaus Schwarz
  12. Peptide Research Group, Clinic for Immunology, Hannover Medical School, Feodor-Lynen Str. 31, 30625 Hannover, Germany

    • Ludger Ständker
    •  & Wolf-Georg Forssmann
  13. A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Vavilova St. 28, 119991 Moscow, Russia

    • Pavel G. Khalatur
  14. Faculty of Physics, Lomonosov Moscow State University, 1-2 Leninskie Gory, 119991 Moscow, Russia

    • Alexei R. Khokhlov
  15. Department of Chemistry, University of Cambridge, Lensfield Road, CB2 1EW Cambridge, UK

    • Tuomas P. J. Knowles

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Contributions

M.Y. generated virus stocks and performed most transductions. C.M. and T.W. assisted in writing the manuscript and were responsible for the structural elucidation of EF-C fibrils. A.K.S., P.G.K. and A.R.K. performed molecular simulations. V.V. and H.G. conducted the ex vivo gene transfer study. C.W.B. and X.S. performed CD analysis. F.A. and D.P. performed coating experiments and measured zeta potentials. O.Z. and J.M. performed Congo Red and ThT assays, S.M.U. was responsible for the analysis of immobilized EF-C by confocal microscopy. D.S. and C.G. performed the fusion assay and flow cytometry assays. P.W. assisted in microscopy. O.L. and T.S. studied the EF-C interaction with virions and cells. J.B. provided retro- and lentiviral vectors and knowhow. H.S. and K.S. isolated and provided human stem cells and performed colony-forming unit assays. L.S. and W.G.F. synthesized and provided peptides. N.R.R. assisted in planning and writing. T.K. analysed AFM data. F.K. and J.M. planned research, analysed data and wrote the manuscript. All authors discussed the results and commented on the manuscript.

Competing interests

M. Yo., F. Ki. and J. Mü. filed for a patent to use EF-C fibrils as tranduction and infection enhancer.

Corresponding author

Correspondence to Jan Münch.

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DOI

https://doi.org/10.1038/nnano.2012.248

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