Random peptide libraries displayed on adeno-associated virus to select for targeted gene therapy vectors.
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.
Müller
Oliver J
OJ
Deutsches Krebsforschungszentrum, Forschungsschwerpunkt Angewandte Tumorvirologie, Im Neuenheimer Feld 242, D-69120 Heidelberg, Germany.
Kaul
Felix
F
Weitzman
Matthew D
MD
Pasqualini
Renata
R
Arap
Wadih
W
Kleinschmidt
Jürgen A
JA
Trepel
Martin
M
D000595:Amino Acid Sequence; D002213:Capsid / Q000378:metabolism; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D018390:Gene Targeting / Q000379:methods*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D019151:Peptide Library; D020539:Sequence Analysis, Protein; D014161:Transduction, Genetic / Q000379:methods; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D019151:Peptide Library; D014764:Viral Proteins