TY - JOUR

T1 - Receptor-Mediated Transfer of DNA-Galactosylated Poly-L-lysine Complexes into Mammalian Cells in vitro and in vivo

AU - Dizhe, E. B.

AU - Akifiev, B. N.

AU - Missul, B. V.

AU - Orlov, S. V.

AU - Kidgotko, O. V.

AU - Sukonina, V. E.

AU - Denisenko, A. D.

AU - Perevozchikov, A. P.

N1 - Funding Information: This work was supported by subprograms “Human Genome” (project No. 88) and “Newest Methods in Bioengineering” (project No. 12) of the direction of prior ity “Technology of Living Systems” of the Scientific Technical Program “Investigations and Elaboration of Civilian Directions of Priority in Science and Technology” of the Russian Federation.

PY - 2001

Y1 - 2001

N2 - With the goal of developing non-viral techniques for exogenous gene delivery into mammalian cells, we have studied receptor-mediated gene transfer using complexes of plasmid DNA and galactosylated poly-L-lysine, poly(L-Lys)Gal. To evaluate the optimal parameters for efficient gene transfer into human hepatoma HepG2 cells by the DNA-poly(L-Lys)Gal complexes, the bacterial reporter genes lacZ and cat were used. Examination of the reporter gene expression level showed that the efficiency of DNA delivery into the cells depends on the structure of DNA-poly(L-Lys)Gal complexes formed at various ionic strength values. The efficiency of DNA transfer into the cells also depends on DNA/poly(L-Lys)Gal molar ratio in the complexes. Plasmid vector carrying human apolipoprotein A-1 (apoA-I) gene was injected as its complex with poly(L-Lys)Gal into rat tail vein. Some level of ApoA-I was detected in the serum of the injected rats. Also, the human apoA-I-containing plasmid was found to be captured specifically by the rat liver cells and transported into the cell nuclei, where it can persist as an episome-like structure for at least a week. After repeated injections of DNA-poly(L-Lys)Gal complexes, the level of human ApoA-I in rat serum increases, probably, due to accumulation of functional human apoA-I gene in the liver cell nuclei. The data seem to be useful for the development of non-viral approaches to gene therapy of cardiovascular diseases.

AB - With the goal of developing non-viral techniques for exogenous gene delivery into mammalian cells, we have studied receptor-mediated gene transfer using complexes of plasmid DNA and galactosylated poly-L-lysine, poly(L-Lys)Gal. To evaluate the optimal parameters for efficient gene transfer into human hepatoma HepG2 cells by the DNA-poly(L-Lys)Gal complexes, the bacterial reporter genes lacZ and cat were used. Examination of the reporter gene expression level showed that the efficiency of DNA delivery into the cells depends on the structure of DNA-poly(L-Lys)Gal complexes formed at various ionic strength values. The efficiency of DNA transfer into the cells also depends on DNA/poly(L-Lys)Gal molar ratio in the complexes. Plasmid vector carrying human apolipoprotein A-1 (apoA-I) gene was injected as its complex with poly(L-Lys)Gal into rat tail vein. Some level of ApoA-I was detected in the serum of the injected rats. Also, the human apoA-I-containing plasmid was found to be captured specifically by the rat liver cells and transported into the cell nuclei, where it can persist as an episome-like structure for at least a week. After repeated injections of DNA-poly(L-Lys)Gal complexes, the level of human ApoA-I in rat serum increases, probably, due to accumulation of functional human apoA-I gene in the liver cell nuclei. The data seem to be useful for the development of non-viral approaches to gene therapy of cardiovascular diseases.

KW - Bacterial reporter genes

KW - Cat gene

KW - Cell nucleus

KW - DNA complex

KW - Episome-like structure

KW - Exogenous dna

KW - Galactosylated poly-L-lysine

KW - Gene delivery

KW - Human apolipoprotein A-I gene

KW - LacZ gene

KW - Mammalian cells

KW - Persistence

UR - http://www.scopus.com/inward/record.url?scp=0035228872&partnerID=8YFLogxK

U2 - 10.1023/A:1002881612906

DO - 10.1023/A:1002881612906

M3 - Article

C2 - 11240393

AN - SCOPUS:0035228872

VL - 66

SP - 55

EP - 61

JO - Biochemistry (Moscow)

JF - Biochemistry (Moscow)

SN - 0006-2979

IS - 1

ER -