An Epidermal Growth Factor-like Repeat of Del1 Protein Increases the Efficiency of Gene Transfer In Vitro

In this paper, we found that Del1, an extracellular matrix protein secreted by embryonic endothelial cells, increases the efficiency of transfection in vitro. Conditioned medium containing Del1 increased transfection by the LacZ gene using several non-viral gene transfer systems, including lipoplex...

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Bibliographic Details
Published in:Molecular biotechnology 2008-07, Vol.39 (3), p.179-185
Main Authors: Kitano, Hisataka, Hidai, Chiaki, Kawana, Masatoshi, Kokubun, Shinichiro
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Biochemistry
Biological Techniques
Biotechnology
Carrier Proteins - physiology
Cell Biology
Cercopithecus aethiops
Chemistry
Chemistry and Materials Science
COS Cells
Culture Media, Conditioned
DNA Primers
Genes
Human Genetics
Humans
In Vitro Techniques
Molecular biology
Protein Science
Proteins
Studies
Transfection - standards
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Summary:In this paper, we found that Del1, an extracellular matrix protein secreted by embryonic endothelial cells, increases the efficiency of transfection in vitro. Conditioned medium containing Del1 increased transfection by the LacZ gene using several non-viral gene transfer systems, including lipoplex and polyplex methods. Experiments using deletion mutants and fragments of Del1 revealed that the third epidermal growth factor-like repeat (E3) of Del1 mediates the enhancement of gene transfer and, furthermore, that the motif CXDXXXFXCXC is essential. Incubation of Pro5 cells, a yolk sac-derived cell line, with as low as 16 pM recombinant E3 was sufficient to enhance transfection, and 1 nM recombinant E3 enhanced the transfection 12-fold. Inhibitors of endocytosis suppressed this activity of the recombinant E3. These results suggest that the E3 fragment of Del1 can be used as a general biological enhancer of non-viral gene transfer.
ISSN:1073-6085
1559-0305
DOI:10.1007/s12033-008-9038-7