Nuclear targeting of non-viral gene carriers using psoralen-nuclear localization signal (NLS) conjugates.
A nuclear localization signal was non-covalently attached to DNA for the purpose of enhancing transfection efficiencies of non-viral gene carriers. Psoralen, a nucleic acid-intercalating agent, was chemically attached to a signal peptide. The conjugate spontaneously intercalated into DNA and then poly(ethyleneimine) [PEI] was added to prepare a DNA/PEI complex containing the signal peptide moieties. The existence of the conjugate did not alter the complexation process between DNA and PEI, which was confirmed by dynamic light scattering. The conjugate was slowly released from the DNA/PEI complex for 24h, while a burst release was examined when the conjugated was added to DNA without PEI. The complex containing a signal peptide moiety increased transfection efficiencies on COS-1 cells, compared to a mutant signal peptide or a control. Cytotoxicity of the conjugate slowly increased as the amount of the conjugate increased, however, the cytotoxic effect of the conjugate was not significant at the effective concentration of the conjugate for transfections. Therefore, the psoralen-nuclear localization signal is expected to be a potent transfection enhancing agent without a covalent modification of transgenes.
Yoo
Hyuk Sang
HS
Department of Biomaterials Engineering, Kangwon National University, Chuncheon, Republic of Korea.
Jeong
Seo Young
SY
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D002522:Chlorocebus aethiops; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D005363:Ficusin / Q000737:chemistry*; D011564:Furocoumarins / Q000737:chemistry; D007097:Imines / Q000737:chemistry; D018613:Microscopy, Confocal; D015394:Molecular Structure; D019913:Nuclear Localization Signals / Q000737:chemistry / Q000235:genetics*; D017319:Photosensitizing Agents / Q000737:chemistry; D011073:Polyamines / Q000494:pharmacology; D011095:Polyethylenes / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry; D014162:Transfection / Q000379:methods*
D011564:Furocoumarins; D007097:Imines; D019913:Nuclear Localization Signals; D017319:Photosensitizing Agents; D011073:Polyamines; D011095:Polyethylenes; D013388:Succinimides; C505405:poly(ethylene imine); C522164:succinimidyl-(4-(psoralen-8-yloxy))butyrate; D004247:DNA; D005363:Ficusin; C001426:N-hydroxysuccinimide