Uptake by rat liver and intracellular fate of plasmid DNA complexed with poly‐l‐lysine or poly‐d‐lysine

FEBS Letters

Laurent, Nathanael¹  Leontieva, Ekaterina¹  Warnier-Pirotte, Marie-Thérèse¹  Jadot, Michel¹  Wattiaux, Robert¹  Wattiaux-De Coninck, Simone¹  Mihaylova, Eugenie¹ 

[1] Laboratoire de Chimie Physiologique, Facultés Universitaires Notre-Dame de la Paix, 61, rue de Bruxelles, B-5000 Namur, Belgium

关键词: Lysosome;    Cationic peptide;    Transfection;    Non-viral vector;   

DOI  :  10.1016/S0014-5793(98)01677-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Efficiency of transfection is probably dependent on the rate of intracellular degradation of plasmid DNA. When a non-viral vector is used, it is not known to what extent the plasmid DNA catabolism is subordinated to the catabolism of the vector. In the work reported here, the problem was approached by following the intracellular fate in rat liver, of plasmid [³⁵S]DNA complexed with a cationic peptide poly-l-lysine that can be hydrolyzed by cellular peptidases or with its stereoisomer, poly-d-lysine, that cannot be split by these enzymes. Complexes of DNA with poly-l-lysine and poly-d-lysine are taken up to the same extent by the liver, mainly by Kupffer cells, but the intracellular degradation of nucleic acid molecules is markedly quicker when poly-l-lysine is injected. The association of DNA with the polycations inhibits DNA hydrolysis in vitro by purified lysosomes but similarly for poly-l-lysine and poly-d-lysine. The intracellular journey followed by [³⁵S]DNA complexed with poly-l- or poly-d-lysine was investigated using differential and isopycnic centrifugation. Results indicate that [³⁵S]DNA is transferred more slowly to lysosomes, the main site of intracellular degradation of endocytosed macromolecules, when it is given as a complex with poly-d-lysine than with poly-l-lysine. They suggest that the digestion of the vector in a prelysosomal compartment is required to allow endocytosed plasmid DNA to rapidly reach lysosomes. Such a phenomenon could explain why injected plasmid DNA is more stable in vivo when it is associated with poly-d-lysine.

【 授权许可】

Unknown