Gene transfer into the lung by nanoparticle dextran-spermine/plasmid DNA complexes.
A novel cationic polymer, dextran-spermine (D-SPM), has been found to mediate gene expression in a wide variety of cell lines and in vivo through systemic delivery. Here, we extended the observations by determining the optimal conditions for gene expression of D-SPM/plasmid DNA (D-SPM/pDNA) in cell lines and in the lungs of BALB/c mice via instillation delivery. In vitro studies showed that D-SPM could partially protect pDNA from degradation by nuclease and exhibited optimal gene transfer efficiency at D-SPM to pDNA weight-mixing ratio of 12. In the lungs of mice, the levels of gene expression generated by D-SPM/pDNA are highly dependent on the weight-mixing ratio of D-SPM to pDNA, amount of pDNA in the complex, and the assay time postdelivery. Readministration of the complex at day 1 following the first dosing showed no significant effect on the retention and duration of gene expression. The study also showed that there was a clear trend of increasing size of the complexes as the amount of pDNA was increased, where the sizes of the D-SPM/pDNA complexes were within the nanometer range.
Abdullah
Syahril
S
UPM-MAKNA Cancer Research Laboratory, Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. syahril@medic.upm.edu.my
Wendy-Yeo
Wai Yeng
WY
Hosseinkhani
Hossein
H
Hosseinkhani
Mohsen
M
Masrawa
Ehab
E
Ramasamy
Rajesh
R
Rosli
Rozita
R
Rahman
Sabariah A
SA
Domb
Abraham J
AJ
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004247:DNA; D003850:Deoxyribonuclease I; D003911:Dextrans / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D005434:Flow Cytometry; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D008156:Luciferases; D008168:Lung / Q000378:metabolism*; D046911:Macromolecular Substances; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D053758:Nanoparticles; D010316:Particle Size; D010957:Plasmids / Q000235:genetics*; D013096:Spermine / Q000737:chemistry*
D003911:Dextrans; D046911:Macromolecular Substances; D013096:Spermine; D004247:DNA; D008156:Luciferases; D003850:Deoxyribonuclease I