Nonviral gene delivery by tetraamino fullerene.
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.
Isobe
Hiroyuki
H
Department of Chemistry, Graduate School of Medicine, The University of Tokyo, Hongo, Tokyo 113-0033, Japan.
Nakanishi
Waka
W
Tomita
Naoki
N
Jinno
Shigeki
S
Okayama
Hiroto
H
Nakamura
Eiichi
E
D000818:Animals; D019556:COS Cells; D002462:Cell Membrane / Q000187:drug effects; D002522:Chlorocebus aethiops; D003571:Cytochalasin B / Q000494:pharmacology; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism*; D016503:Drug Delivery Systems / Q000379:methods*; D004587:Electrophoresis, Agar Gel; D037741:Fullerenes / Q000737:chemistry* / Q000494:pharmacology*; D005822:Genetic Vectors; D015394:Molecular Structure; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
D004279:DNA, Viral; D037741:Fullerenes; D012367:RNA, Viral; C510698:tetraamino fullerene; D003571:Cytochalasin B