Isolation of targeted AAV2 vectors from novel virus display libraries
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Isolation of targeted AAV2 vectors from novel virus display libraries. / Waterkamp, Daniel A; Müller, Oliver J; Ying, Ying; Trepel, Martin; Kleinschmidt, Jürgen A.
In: J GENE MED, Vol. 8, No. 11, 11.2006, p. 1307-19.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Isolation of targeted AAV2 vectors from novel virus display libraries
AU - Waterkamp, Daniel A
AU - Müller, Oliver J
AU - Ying, Ying
AU - Trepel, Martin
AU - Kleinschmidt, Jürgen A
PY - 2006/11
Y1 - 2006/11
N2 - Random peptide ligands displayed on viral capsids are emerging tools for selection of targeted gene transfer vectors even without prior knowledge of the potential target cell receptor. We have previously introduced adeno-associated viral (AAV)-displayed peptide libraries that ensure encoding of displayed peptides by the packaged AAV genome. A major limitation of these libraries is their contamination with wild-type (wt) AAV. Here we describe a novel and improved library production system that reliably avoids generation of wt AAV by use of a synthetic cap gene. Selection of targeted AAV vectors from wt-containing and the novel wt-free libraries on cell types with different permissivity for wt AAV2 replication suggested the superiority of the wt-free library. However, from both libraries highly specific peptide sequence motifs were selected which improved transduction of cells with moderate or low permissivity for AAV2 replication. Strong reduction of HeLa cell transduction compared to wt AAV2 and only low level transduction of non-target cells by some selected clones showed that not only the efficiency but also the specificity of gene transfer was improved. In conclusion, our study validates and improves the unique potential of virus display libraries for the development of targeted gene transfer vectors.
AB - Random peptide ligands displayed on viral capsids are emerging tools for selection of targeted gene transfer vectors even without prior knowledge of the potential target cell receptor. We have previously introduced adeno-associated viral (AAV)-displayed peptide libraries that ensure encoding of displayed peptides by the packaged AAV genome. A major limitation of these libraries is their contamination with wild-type (wt) AAV. Here we describe a novel and improved library production system that reliably avoids generation of wt AAV by use of a synthetic cap gene. Selection of targeted AAV vectors from wt-containing and the novel wt-free libraries on cell types with different permissivity for wt AAV2 replication suggested the superiority of the wt-free library. However, from both libraries highly specific peptide sequence motifs were selected which improved transduction of cells with moderate or low permissivity for AAV2 replication. Strong reduction of HeLa cell transduction compared to wt AAV2 and only low level transduction of non-target cells by some selected clones showed that not only the efficiency but also the specificity of gene transfer was improved. In conclusion, our study validates and improves the unique potential of virus display libraries for the development of targeted gene transfer vectors.
KW - Animals
KW - Base Sequence
KW - Cell Line
KW - DNA Primers
KW - Dependovirus
KW - Gene Transfer Techniques
KW - Genetic Therapy
KW - Genetic Vectors
KW - HeLa Cells
KW - Humans
KW - Luciferases
KW - Mice
KW - Peptide Library
KW - Plasmids
KW - Rats
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1002/jgm.967
DO - 10.1002/jgm.967
M3 - SCORING: Journal article
C2 - 16955542
VL - 8
SP - 1307
EP - 1319
JO - J GENE MED
JF - J GENE MED
SN - 1099-498X
IS - 11
ER -
