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 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.
Kitano
Hisataka
H
Department of Dental Surgery, Nihon University School of Medicine, Tokyo 173-8610, Japan.
Hidai
Chiaki
C
Kawana
Masatoshi
M
Kokubun
Shinichiro
S
D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins / Q000502:physiology*; D015815:Cell Adhesion Molecules; D002522:Chlorocebus aethiops; D017077:Culture Media, Conditioned; D017931:DNA Primers; D006801:Humans; D066298:In Vitro Techniques; D014162:Transfection / Q000592:standards*
D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D017077:Culture Media, Conditioned; D017931:DNA Primers; C110183:EDIL3 protein, human