The transmembrane domain of diphtheria toxin improves molecular conjugate gene transfer

被引:47
作者
Fisher, KJ
Wilson, JM
机构
[1] UNIV PENN HLTH SYST, INST HUMAN GENE THERAPY, PHILADELPHIA, PA USA
[2] UNIV PENN HLTH SYST, DEPT MOL & CELLULAR ENGN, PHILADELPHIA, PA USA
[3] WISTAR INST ANAT & BIOL, PHILADELPHIA, PA 19104 USA
关键词
D O I
10.1042/bj3210049
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Vectors based on the formation of a soluble DNA-polycation complex are being developed for the treatment of human diseases. These complexes are rapidly taken up by receptor-mediated endocytosis, but are inefficiently delivered to the nucleus owing to entrapment in membrane-bound vesicles. In this study we introduced the transmembrane domain of diphtheria toxin into a DNA-polycation conjugate complex in an effort to increase gene transfer by membrane perturbation. The transmembrane domain of diphtheria toxin was expressed in Escherichia coli as a maltose-binding protein fusion and chemically coupled to high-molecular-mass poly-L-lysine. Incorporation of this conjugate into a traditional complex formed with a luciferase-containing plasmid with an asialo-orosomucoid-polycation conjugate significantly increased transfection efficiency in vitro in a manner proportional to the amount of diphtheria toxin incorporated. The delivery of luciferase RNA transcript was similarly increased when complexed with similar polycation conjugates. This study uses the structural biology of a bacterial protein to improve polycation-based gene delivery.
引用
收藏
页码:49 / 58
页数:10
相关论文
共 48 条
[1]  
BASHFORD CL, 1986, J BIOL CHEM, V261, P9300
[2]   REFINED STRUCTURE OF MONOMERIC DIPHTHERIA-TOXIN AT 2.3-ANGSTROM RESOLUTION [J].
BENNETT, MJ ;
EISENBERG, D .
PROTEIN SCIENCE, 1994, 3 (09) :1464-1475
[3]   EFFECT OF PH ON THE CONFORMATION OF DIPHTHERIA-TOXIN AND ITS IMPLICATIONS FOR MEMBRANE PENETRATION [J].
BLEWITT, MG ;
CHUNG, LA ;
LONDON, E .
BIOCHEMISTRY, 1985, 24 (20) :5458-5464
[4]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[5]   TARGETED DELIVERY OF ANTISENSE OLIGONUCLEOTIDES BY MOLECULAR CONJUGATES [J].
BUNNELL, BA ;
ASKARI, FK ;
WILSON, JM .
SOMATIC CELL AND MOLECULAR GENETICS, 1992, 18 (06) :559-569
[6]   COMPLEXES OF POLYLYSINE WITH POLYURIDYLIC ACID AND OTHER POLYNUCLEOTIDES [J].
CARROLL, D .
BIOCHEMISTRY, 1972, 11 (03) :426-+
[7]   THE CRYSTAL-STRUCTURE OF DIPHTHERIA-TOXIN [J].
CHOE, S ;
BENNETT, MJ ;
FUJII, G ;
CURMI, PMG ;
KANTARDJIEFF, KA ;
COLLIER, RJ ;
EISENBERG, D .
NATURE, 1992, 357 (6375) :216-222
[8]  
CHOWDHURY NR, 1993, J BIOL CHEM, V268, P11265
[9]   CONFORMATIONAL ANALYSIS OF DNA-POLY-L-LYSINE COMPLEXES BY OPTICAL ROTATORY DISPERSION [J].
COHEN, P ;
KIDSON, C .
JOURNAL OF MOLECULAR BIOLOGY, 1968, 35 (01) :241-&
[10]   HIGH-EFFICIENCY RECEPTOR-MEDIATED DELIVERY OF SMALL AND LARGE (48 KILOBASE GENE CONSTRUCTS USING THE ENDOSOME-DISRUPTION ACTIVITY OF DEFECTIVE OR CHEMICALLY INACTIVATED ADENOVIRUS PARTICLES [J].
COTTEN, M ;
WAGNER, E ;
ZATLOUKAL, K ;
PHILLIPS, S ;
CURIEL, DT ;
BIRNSTIEL, ML .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1992, 89 (13) :6094-6098