Dexamethasone conjugated poly(amidoamine) dendrimer as a gene carrier for efficient nuclear translocation.
Nuclear membrane is one of the main barriers in polymer-mediated intracellular gene delivery. It was previously reported that glucocorticoid receptor dilated the nuclear pore and translocated into nucleus when it bound to its ligand, glucocorticoid. This suggests that the transport of DNA into nucleus may be facilitated by glucocorticoid. In this study, a glucocorticoid, dexamethasone, was conjugated to polyamidoamine (PAMAM) dendrimer and the effect of the conjugation was investigated. The PAMAM-Dexamethasone (PAM-Dexa) was synthesized by the one-step reaction using Traut's reagent. PAM-Dexa/plasmid DNA complex was completely retarded at a 1/1 weight ratio (polymer/DNA) in a gel retardation assay. PAM-Dexa protected DNA from DNase I for more than 60 min. PAM-Dexa/plasmid DNA complex showed the highest transfection efficiency to 293 cells at a 0.8/1 weight ratio. At this ratio, PAM-Dexa had higher transfection efficiency than PAMAM. Especially in the presence of serum during the transfection, the transfection efficiency of PAM-Dexa was higher than that of PAMAM or PEI by one order of magnitude. In addition, more PAM-Dexa/DNA complexes were observed in the nucleus region than PAMAM/DNA from the confocal microscopy studies. These results indicated that the technique with dexamethasone might be useful for the gene delivery using polymeric gene carriers and the development of efficient polymer vectors.
Choi
Joon Sig
JS
Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon 305-764, South Korea.
Ko
Kyung Soo
KS
Park
Jong Sang
JS
Kim
Yong-Hee
YH
Kim
Sung Wan
SW
Lee
Minhyung
M
D021581:Active Transport, Cell Nucleus; D000818:Animals; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D050091:Dendrimers; D003850:Deoxyribonuclease I; D003907:Dexamethasone / Q000737:chemistry* / Q000633:toxicity; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D051379:Mice; D018613:Microscopy, Confocal; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D010316:Particle Size; D011073:Polyamines / Q000737:chemistry* / Q000633:toxicity; D014162:Transfection
D050091:Dendrimers; C104700:PAMAM Starburst; D011073:Polyamines; D003907:Dexamethasone; D004247:DNA; D008156:Luciferases; D003850:Deoxyribonuclease I