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Article
Nature Biotechnology  21, 1040 - 1046 (2003)
Published online: 3 August 2003; Corrected online: 17 August 2003 | doi:10.1038/nbt856

Random peptide libraries displayed on adeno-associated virus to select for targeted gene therapy vectors

Oliver J Müller1, 6, Felix Kaul2, 6, Matthew D Weitzman4, Renata Pasqualini5, Wadih Arap5, Jürgen A Kleinschmidt1, 6 & Martin Trepel2, 3, 6

1  Deutsches Krebsforschungszentrum, Forschungsschwerpunkt Angewandte Tumorvirologie, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany.

2  University of Freiburg Medical Center, Department of Hematology and Oncology, Hugstetter Strasse 55, D-79106 Freiburg, Germany.

3  University of Freiburg Medical Center, Institute for Molecular Medicine and Cell Research, Hugstetter Strasse 55, D-79106 Freiburg, Germany.

4  The Salk Institute, Laboratory of Genetics, P.O. Box 85800, La Jolla, California 92183-5800, USA.

5  The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Blvd., Unit 427, Houston, Texas 77030-4095, USA.

6  These authors contributed equally to this work.

Correspondence should be addressed to Jürgen A Kleinschmidt j.kleinschmidt@dkfz-heidelberg.de
Characterizing the molecular diversity of the cell surface is critical for targeting gene therapy. Cell type−specific binding ligands can be used to target gene therapy vectors. However, targeting systems in which optimum eukaryotic vectors can be selected on the cells of interest are not available. Here, we introduce and validate a random adeno-associated virus (AAV) peptide library in which each virus particle displays a random peptide at the capsid surface. This library was generated in a three-step system that ensures encoding of displayed peptides by the packaged DNA. As proof-of-concept, we screened AAV-libraries on human coronary artery endothelial cells. We observed selection of particular peptide motifs. The selected peptides enhanced transduction in coronary endothelial cells but not in control nonendothelial cells. This vector targeting strategy has advantages over other combinatorial approaches such as phage display because selection occurs within the context of the capsid and may have a broad range of applications in biotechnology and medicine.

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REFERENCE
Adeno-associated Viruses
Nature Encyclopaedia of Life Sciences

REVIEWS
Progress and problems with the use of viral vectors for gene therapy
Nature Reviews Genetics Review (01 May 2003)
Viral vectors for gene therapy: the art of turning infectious agents into vehicles of therapeutics
Nature Medicine Review Article (01 Jan 2001)

NEWS AND VIEWS
AAV display—homing in on the target
Nature Biotechnology News and Views (01 Sep 2003)

RESEARCH
Genetic capsid modifications allow efficient re-targeting of adeno-associated virus type 2
Nature Medicine Article (01 Sep 1999)

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Nature Biotechnology
ISSN: 1087-0156
EISSN: 1546-1696
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