Poly(L-lactide-co-glycolide) nanospheres conjugated with a nuclear localization signal for delivery of plasmid DNA.
Polymeric nanospheres fabricated from biodegradable poly(lactide-co-glycolide) (PLGA) have been extensively investigated for applications in gene delivery. In this study, we show that the covalent conjugation of a nuclear localization signal (NLS, SV40 peptide) on PLGA nanospheres enhances the gene transfection efficiency. NLS conjugated PLGA copolymer was prepared by using a coupling reaction between maleimide-terminated PLGA copolymer and NLS in the presence of Imject maleimide conjugation buffer. PLGA nanospheres encapsulating plasmid (pDNA) were prepared by using a double emulsion-solvent evaporation method. The kinetics of in vitro release of pDNA from PLGA nanospheres was determined with UV in phosphate buffered saline (PBS). Gene transfection efficiency in human dermal fibroblasts was tested in vitro using nanospheres encapsulating the luciferase gene. The conjugation of the NLS peptide to the PLGA nanospheres could improve the nuclear localization and/or cellular uptake of PLGA nanosphere/pDNA constructs and thereby improve the transfection efficiency of a PLGA nanosphere gene delivery system. The pDNA was released from PLGA nanospheres over nine days. NLS conjugation enhanced the gene transfection efficiency in vitro by 1.2 approximately 3.2-fold over 13 days. PLGA/pDNA nanospheres appeared to be superior to PEI/pDNA complexes for the long-term expression of pDNA. Furthermore, the level of the sustained gene expression of the PLGA nanospheres was enhanced by the conjugation of NLS to the PLGA nanospheres. This study showed that the NLS conjugation enhanced the gene transfection efficiency of the PLGA nanosphere gene delivery system in vitro and that the enhanced gene expression was sustained for at least 13 days.
Jeon
Oju
O
Departments of Chemical Engineering, Hanyang University, Seoul, South Korea.
Lim
Hee-Won
HW
Lee
Minhyung
M
Song
Su Jin
SJ
Kim
Byung-Soo
BS
D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D018014:Gene Transfer Techniques; D006801:Humans; D019344:Lactic Acid / Q000009:adverse effects / Q000737:chemistry*; D043942:Nanotubes / Q000009:adverse effects / Q000737:chemistry*; D019913:Nuclear Localization Signals / Q000009:adverse effects / Q000737:chemistry*; D010316:Particle Size; D010957:Plasmids / Q000008:administration & dosage* / Q000235:genetics; D011100:Polyglycolic Acid / Q000009:adverse effects / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers / Q000009:adverse effects / Q000737:chemistry*; D014162:Transfection
D019913:Nuclear Localization Signals; D011108:Polymers; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid; D004247:DNA