Nonviral Gene Delivery by Tetraamino Fullerene

• Published in: Molecular pharmaceutics 2006-03, Vol.3 (2), p.124-134
• Main Authors: Isobe, Hiroyuki, Nakanishi, Waka, Tomita, Naoki, Jinno, Shigeki, Okayama, Hiroto, Nakamura, Eiichi
• Format: Article
• Language: English
• Subjects: Animals; Cell Membrane - drug effects; Cercopithecus aethiops; COS Cells; Cytochalasin B - pharmacology; DNA, Viral - genetics; DNA, Viral - metabolism; Drug Delivery Systems - methods; Electrophoresis, Agar Gel; Fullerenes - chemistry; Fullerenes - pharmacology; Genetic Vectors; Molecular Structure; RNA, Viral - genetics; RNA, Viral - metabolism; Transfection - methods
• ISSN: 1543-8384; 1543-8392
• DOI: 10.1021/mp050068r

A fullerene derivative bearing two diamino side chains binds to a plasmid vector DNA, either 4 or 40 kbp in size, delivers it to mammalian cells on incubation, and leads to expression of the encoded gene either transiently or stably. The initial physicochemical investigations upon DNA-binding and protective properties of various fullerene compounds against nuclease led us to identify the tetraamino fullerene as an ideal candidate to probe the new concept of fullerene-mediated gene delivery to mammalian cells. Studies on transient and stable transfection of COS-1 cells using green fluorescent protein and luciferase reporter genes revealed several useful properties of the fullerene transfection as compared with the conventional lipid-based transfection method, including much higher efficiency of stable transfection and ability to transfect confluent cells. Chemical and biological studies suggested that the cell uptake of the fullerene/DNA complex takes place by an endocytosis mechanism and that the DNA internalized by endosomes is protected by the fullerene against enzymatic digestion. The stiffness of the fullerene/DNA complex may play some role in the success of the fullerene method. Keywords: Fullerene; stable transfection; DNA protection; endocytosis