DNA plasmids formed particulate complexes wit
h a variety of cationic polyamino acids and cationic lipids, w
hic
h were used to transfect mammalian cells in culture. Complexation was studied by assaying for exclusion of et
hidium using a fluorometric assay, w
hic
h indicated t
hat complexation wit
h cationic polyamino acids took place wit
h utilisation of t
he majority of c
harged functional groups. T
he particle sizes and zeta potentials of a range of complexes were determined. Generally polyamino acids formed uniform particles 80&ndas
h;120 nm in diameter in water, but t
heir particle size increased on dilution of t
he particles in electrolytes or cell culture media. T
he efficiency of transfection was compared using complexes of pRSV
lacZ, a reporter construct w
hic
h expressed β-galactosidase under t
he control of t
he Rous sarcoma virus promoter. Positively c
harged DNA/polyamino acid complexes were taken up by cells but required an endosomolytic agent, suc
h as c
hloroquine, to facilitate transfection. Polyornit
hine complexes resulted in t
he
hig
hest levels of expression, in comparison wit
h ot
her
homopolyamino acids (polyornit
hine>poly-l-lysine=poly-d-lysine>polyarginine). Copolyamino acids of lysine and alanine condensed DNA but were less active in transfection experiments. Copoly(l-Lys, l-Ala 1:1) was inactive even in t
he presence of c
hloroquine. In contrast DNA/cationic lipid complexes transfected cells spontaneously, and c
hloroquine did not improve t
he extent of expression, rat
her it usually reduced efficiency. T
here was little correlation between comparative efficiencies of lipid complexes between cell lines suggesting t
hat t
he nature of t
he cell membrane and differences in mec
hanisms of internalisation were determinants of efficiency. In an effort to explore better cell culture models for gene delivery, monolayers of Caco-2 cells were transfected in filter culture. As t
he cells differentiated and formed a polarized monolayer, expression of β-galactosidase was reduced until at day 27 expression was not significantly different from basal activity. T
he Caco-2 filter culture model merits furt
her attention as a model of gene delivery to epit
helial surfaces, suc
h as would be encountered in t
he lung after in
halation.
h=""100"" bgcolor=""#F0F0F0"" valign=""top""> Publisher: | h=""470"" valign=""top""> Elsevier Science |
h=""100"" bgcolor=""#F0F0F0"" valign=""top""> Language of Publication: | h=""470"" valign=""top""> English |
h=""100"" bgcolor=""#F0F0F0"" valign=""top""> Item Identifier: | h=""470"" valign=""top""> S0168-3659(98)00015-7 |
h=""100"" bgcolor=""#F0F0F0"" valign=""top""> Publication Type: | h=""470"" valign=""top""> Article |
h=""100"" bgcolor=""#F0F0F0"" valign=""top""> ISSN: | h=""470"" valign=""top""> 0168-3659 |
h=""100"" bgcolor=""#F0F0F0"" valign=""top""> Cited by: | h=""470"" valign=""top""> - Brown, Maureen D.; Schatzlein, Andreas G.; Uchegbu, Ijeoma F.,""Gene delivery with synthetic (non viral) carriers""International Journal of Pharmaceutics2001pp. 1-21
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Seymour, Leonard W.,""Key issues in non-viral gene deliveryhref=""#footnote_1"">1""Advanced Drug Delivery Reviews2001pp. 187-203 href=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=0169409x&issue=v46i1-3&article=187_kiingd"">Bibliographic Pagehref=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=0169409x&issue=v46i1-3&article=187_kiingd&form=pdf&file=file.pdf"">Full TextLee, Haeshin; Jeong, Ji Hoon; Park, Tae Gwan,""PEG grafted polylysine with fusogenic peptide for gene delivery: high transfection efficiency with low cytotoxicity""Journal of Controlled Release2002pp. 283-291 href=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=01683659&issue=v79i1-3&article=283_pgpwfphtewlc"">Bibliographic Pagehref=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=01683659&issue=v79i1-3&article=283_pgpwfphtewlc&form=pdf&file=file.pdf"">Full TextLu, Yingjuan; Low, Philip S.,""Folate-mediated delivery of macromolecular anticancer therapeutic agents""Advanced Drug Delivery Reviews2002pp. 675-693 href=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=0169409x&issue=v54i0005&article=675_fdomata"">Bibliographic Pagehref=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=0169409x&issue=v54i0005&article=675_fdomata&form=pdf&file=file.pdf"">Full TextMerdan, Thomas; 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Evensen, Oystein; Thu, Beate J.,""The influence of formulation variables on in vitro transfection efficiency and physicochemical properties of chitosan-based polyplexes""International Journal of Pharmaceutics2003pp. 115-127 href=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=03785173&issue=v261i1-2&article=115_tiofvoappocp"">Bibliographic Pagehref=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=03785173&issue=v261i1-2&article=115_tiofvoappocp&form=pdf&file=file.pdf"">Full TextChamarthy, Sai Prasanth; Kovacs, Jeffrey R.; McClelland, Erin; Gattens, Diane; Meng, Wilson S.,""A cationic peptide consists of ornithine and histidine repeats augments gene transfer in dendritic cells""Molecular Immunology2003pp. 483-490 href=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=01615890&issue=v40i0008&article=483_acpcooagtidc"">Bibliographic Pagehref=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=01615890&issue=v40i0008&article=483_acpcooagtidc&form=pdf&file=file.pdf"">Full TextBrown, Maureen D.; Gray, Alexander I.; Tetley, Laurence; Santovena, Ana; Rene, Judith; Schatzlein, Andreas G.; Uchegbu, Ijeoma F.,""In vitro and in vivo gene transfer with poly(amino acid) vesicles""Journal of Controlled Release2003pp. 193-211 href=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=01683659&issue=v93i0002&article=193_ivaivgtwpav"">Bibliographic Pagehref=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=01683659&issue=v93i0002&article=193_ivaivgtwpav&form=pdf&file=file.pdf"">Full TextEl-Aneed, Anas,""An overview of current delivery systems in cancer gene therapy""Journal of Controlled Release2004pp. 1-14 href=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=01683659&issue=v94i0001&article=1_aoocdsicgt"">Bibliographic Pagehref=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=01683659&issue=v94i0001&article=1_aoocdsicgt&form=pdf&file=file.pdf"">Full TextTokunaga, Masaki; Hazemoto, Norio; Yotsuyanagi, Toshihisa,""Effect of oligopeptides on gene expression: comparison of DNA/peptide and DNA/peptide/liposome complexes""International Journal of Pharmaceutics2004pp. 71-80 href=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=03785173&issue=v269i0001&article=71_eooogecodadc"">Bibliographic Pagehref=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=03785173&issue=v269i0001&article=71_eooogecodadc&form=pdf&file=file.pdf"">Full TextRamsay, Euan; Hadgraft, Jon; Birchall, James; Gumbleton, Mark,""Examination of the biophysical interaction between plasmid DNA and the polycations, polylysine and polyornithine, as a basis for their differential gene transfection in-vitro""International Journal of Pharmaceutics2000pp. 97-107 href=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=03785173&issue=v210i1-2&article=97_eotbibftdgti"">Bibliographic Pagehref=""http://202.120.13.78/cgi-bin/sciserv.pl?collection=journals&journal=03785173&issue=v210i1-2&article=97_eotbibftdgti&form=pdf&file=file.pdf"">Full Text |
h=""30""> | <hr size=""1""/> Footnotes: PII of original article: S0169-409X(98)00048-9. The article was originally published in Advanced Drug Delivery Reviews 34 (1998) 3-19.
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