Abstract

We present a flexible and highly specific targeting method for lentiviral vectors based on single-chain antibodies recognizing cell-surface antigens. We generated lentiviral vectors specific for human CD105+ endothelial cells, human CD133+ hematopoietic progenitors and mouse GluA-expressing neurons. Lentiviral vectors specific for CD105 or for CD20 transduced their target cells as efficiently as VSV-G pseudotyped vectors but discriminated between endothelial cells and lymphocytes in mixed cultures. CD133-targeted vectors transduced CD133+ cultured hematopoietic progenitor cells more efficiently than VSV-G pseudotyped vectors, resulting in stable long-term transduction. Lentiviral vectors targeted to the glutamate receptor subunits GluA2 and GluA4 exhibited more than 94% specificity for neurons in cerebellar cultures and when injected into the adult mouse brain. We observed neuron-specific gene modification upon transfer of the Cre recombinase gene into the hippocampus of reporter mice. This approach allowed targeted gene transfer to many cell types of interest with an unprecedented degree of specificity.

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Acknowledgements

This work was supported by grants from the Priority Programme “Mechanisms of gene vector entry and persistence” of the Deutsche Forschungsgemeinschaft to C.J.B. and K.C. and from the 7th European Community programme project Persisting Transgenesis (Persist) to C.J.B. U.C.M. was supported by grants of the Deutsche Forschungsgemeinschaft (MU 1457/8-1) and the “Nationales Genomforschungsnetz” (01GS08128). T.A. is supported by the graduate study program GK1172 Biologicals of the Goethe University Frankfurt AM.

Author information

Author notes

    • Brigitte Anliker
    • , Tobias Abel
    •  & Sabrina Kneissl

    These authors contributed equally to this work.

Affiliations

  1. Division of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.

    • Brigitte Anliker
    • , Tobias Abel
    • , Sabrina Kneissl
    • , Julia Brynza
    • , Irene C Schneider
    • , Robert C Münch
    • , Klaus Cichutek
    •  & Christian J Buchholz
  2. Institute of Virology, Department of Pathobiology, University of Veterinary Medicine, Vienna, Austria.

    • Juraj Hlavaty
    • , Helga Petznek
    •  & Christine Hohenadl
  3. Institute for Clinical Neurobiology, University Hospital of Neurology, Heidelberg, Germany.

    • Antonio Caputi
    •  & Hannah Monyer
  4. Institute of Cell Biology and Immunology, University of Stuttgart, Stuttgart, Germany.

    • Roland E Kontermann
  5. Pediatric Hematology and Oncology, University Hospital, Frankfurt, Germany.

    • Ulrike Koehl
  6. Miltenyi Biotec GmbH, Bergisch-Gladbach, Germany.

    • Ian C D Johnston
  7. Department of Biosciences, Division of Biochemistry, University of Helsinki, Finland.

    • Kari Keinänen
  8. Institute for Pharmacy and Molecular Biotechnology, University of Heidelberg, Germany.

    • Ulrike C Müller

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Contributions

B.A., T.A. and S.K. designed and performed experiments and contributed to writing of the manuscript. J.H., A.C., J.B., I.C.S., R.C.M. and H.P. performed experiments. R.E.K., U.K., I.C.D.J. and K.K. contributed protocols and reagents. C.H. and H.M. supervised work. U.C.M. supervised work and contributed to writing of the manuscript. K.C. acquired grants. C.J.B. conceived and designed the study, acquired grants, supervised work and wrote the manuscript.

Competing interests

I.C.D.J. is an employee of Miltenyi Biotec GmbH. S.K., K.C. and C.J.B. are listed as inventors in an international Patent Cooperation Treaty European patent application (PCT/EP2007/008384) assigned to the Paul Ehrlich Institut, which includes as claims the generation of targeted lentiviral vectors.

Corresponding author

Correspondence to Christian J Buchholz.

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DOI

https://doi.org/10.1038/nmeth.1514

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