Dendritic alpha,epsilon-poly(L-lysine)s as delivery agents for antisense oligonucleotides.
OBJECTIVE
To evaluate the potential use of dendritic alpha,epsilon-poly(L-lysine)s (DPL) for the efficient cellular delivery of antisense oligonucleotides.


METHODS
A series of dendritic alpha,epsilon-polylysines of various generations were prepared. Their physical properties and the ability to form complex with oligonucleotide were investigated by polyacrylamide gel electrophoresis, capillary zone electrophoresis (CZE), agarose gel electrophoresis, fluorescence titration and atomic force microscopy (AFM). The efficiency to deliver oligonucleotide to HeLa cells, stably transfected with plasmid pLuc/705, was evaluated by using antisense splicing correction assay and confocal microscopy.


RESULTS
DPLs formed the complexes with antisense oligonucleotide with modest cytotoxicity. The charge ratio of oligonucleotide to DPL and the size (generation) of DPLs were all critical variables for the antisense effect. Compared to low generation DPLs, high generation DPLs were more effective in delivering oligonucleotide into cells.


CONCLUSIONS
High generation DPL-oligonucleotide complexes were moderately effective for delivery antisense oligonucleotide. The complex formation provides a promise for in vivo therapeutic application of DPLs or their derivatives in the delivery of gene or oligonucleotide.
Eom
Khee Dong
KD
Nanormics Inc., Sungbuk-gu, Seoul, 136-130, South Korea.
Park
Sun Mi
SM
Tran
Huu Dung
HD
Kim
Myong Soo
MS
Yu
Ri Na
RN
Yoo
Hoon
H
D002470:Cell Survival / Q000187:drug effects; D050091:Dendrimers / Q000138:chemical synthesis / Q000737:chemistry*; D004586:Electrophoresis; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D046911:Macromolecular Substances / Q000138:chemical synthesis / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D016376:Oligonucleotides, Antisense / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D011073:Polyamines / Q000737:chemistry; D011107:Polylysine / Q000138:chemical synthesis / Q000737:chemistry*; D012326:RNA Splicing; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014162:Transfection / Q000379:methods
D050091:Dendrimers; D046911:Macromolecular Substances; D016376:Oligonucleotides, Antisense; C104700:PAMAM Starburst; D011073:Polyamines; D011107:Polylysine; D008156:Luciferases