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ISSN 0582-9879 Acta Biochimica et Biophysica Sinica 2004, 36(7): 485–491 CN 31-1300/Q
A Novel Gene Delivery System Targeting Urokinase Receptor
Xing-Hui SUN, Li TAN, Chun-Yang LI, Chang TONG, Jin FAN, Ping LI,
and Yun-Song ZHU*
Department of
Molecular Genetics, Key Laboratory of Molecular Medicine of Ministry Education,
Shanghai Medical College of Fudan University, Shanghai 200032, China
Abstract Recombinant proteins
that combine different functions required for cell targeting and intra-cellular
delivery of DNA present an attractive approach for the development of nonviral
gene delivery vectors. Here, we described a novel protein termed ATF-lys10
which facilitated cell-specific gene transfer via receptor-mediated
endocytosis. ATF-lys10 was composed of the amino-terminal fragment of urokinase
and ten lysines at the carboxyl terminus. Bacterially expressed ATF-lys10
protein existed in soluble form, and had antigenicity of human urokinase.
Purified ATF-lys10 specifically bound to uPAR-expressing cells and formed
protein-DNA complexes with plasmid pGL3-control. After neutralization of excess
negative charge with poly-L-lysine, these complexes served as a specific
gene delivery vector for uPAR-expressing cells. Lyso-somotropic compounds, such
as chloroquine, drastically increased the ATF-lys10 mediated gene delivery
efficiency. Our results suggest that the recombinant protein ATF-lys10 with the
properties of DNA binding and tumor cell targeting represents a promising
method for gene transfer and expression in tumor cells.
Key words amino-terminal fragment;
urokinase; gene transfer; endocytosis; urokinase receptor
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Received: April 5, 2004 Accepted: May 7, 2004
This work was supported by a grant from the National Natural Science
Foundation of China (No. 30170398)
*Corresponding author: Tel, 86-21-54237278; Fax,
86-21-64033738; E-mail, [email protected]