Abstract
Background: Despite the use of currently optimized lipofection conditions, including transfection in serum-depleted media, the efficiency of gene transfer is low and high transfection rates often induce cytotoxicity. A lipid formulation with transfection efficiency not inhibited by serum would provide an advance towards in vivo applications. Methods: We explored the ability of the cationic lipid SH-14 to dimerize upon DNA and form lipoplexes, and potentially release nucleic acids in the intracellular reducing milieu. We investigated the critical micelle-forming concentration of SH-14 and its intrinsic toxicity, size and Zeta potential measurements, the in vitro cytotoxicity of SH-14/ plasmid DNA lipoplexes and their ability to transfect cells. Results: Among all the charge ratios (CR, +/-) tested, lipoplexes at CR 10 with a mean diameter of 295 nm and a surface charge of +20 mV, exhibited the best compromise between transfection efficiency and tolerability. SH-14 presented the same cytotoxicity level whether alone or complexed in lipoplexes. Lipofections carried out in serum-free medium shared a transfection efficiency, on average, of 40% and a cytotoxicity of 38%. An increase of 73% in transfection efficiency and 24% in cell viability were obtained, extending lipofection over 48 h in complete-medium. Moreover, when serum concentration was increased from 10% to 50%, a three-fold increase in plasmid dose led to more than 72% of cells being transfected with almost no sign of cytotoxicity. Conclusions: Overall, SH-14 presents good potential as a novel transfection reagent to be used in the presence of serum.
| Original language | English |
|---|---|
| Pages (from-to) | 637-645 |
| Number of pages | 9 |
| Journal | Journal of Gene Medicine |
| Volume | 10 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Jun 2008 |
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Keywords
- Dimerizable function
- Gene delivery
- Lipoplex
- Nonviral vector
- Serum effect
- Transfection
ASJC Scopus subject areas
- Genetics
Cite this
A dimerizable cationic lipid with potential for gene delivery. / Candiani, Gabriele; Pezzoli, Daniele; Cabras, Mariasara; Ristori, Sandra; Pellegrini, Cinzia; Kajaste-Rudnitski, Anna; Vicenzi, Elisa; Sala, Carlo; Zanda, Matteo.
In: Journal of Gene Medicine, Vol. 10, No. 6, 06.2008, p. 637-645.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A dimerizable cationic lipid with potential for gene delivery
AU - Candiani, Gabriele
AU - Pezzoli, Daniele
AU - Cabras, Mariasara
AU - Ristori, Sandra
AU - Pellegrini, Cinzia
AU - Kajaste-Rudnitski, Anna
AU - Vicenzi, Elisa
AU - Sala, Carlo
AU - Zanda, Matteo
PY - 2008/6
Y1 - 2008/6
N2 - Background: Despite the use of currently optimized lipofection conditions, including transfection in serum-depleted media, the efficiency of gene transfer is low and high transfection rates often induce cytotoxicity. A lipid formulation with transfection efficiency not inhibited by serum would provide an advance towards in vivo applications. Methods: We explored the ability of the cationic lipid SH-14 to dimerize upon DNA and form lipoplexes, and potentially release nucleic acids in the intracellular reducing milieu. We investigated the critical micelle-forming concentration of SH-14 and its intrinsic toxicity, size and Zeta potential measurements, the in vitro cytotoxicity of SH-14/ plasmid DNA lipoplexes and their ability to transfect cells. Results: Among all the charge ratios (CR, +/-) tested, lipoplexes at CR 10 with a mean diameter of 295 nm and a surface charge of +20 mV, exhibited the best compromise between transfection efficiency and tolerability. SH-14 presented the same cytotoxicity level whether alone or complexed in lipoplexes. Lipofections carried out in serum-free medium shared a transfection efficiency, on average, of 40% and a cytotoxicity of 38%. An increase of 73% in transfection efficiency and 24% in cell viability were obtained, extending lipofection over 48 h in complete-medium. Moreover, when serum concentration was increased from 10% to 50%, a three-fold increase in plasmid dose led to more than 72% of cells being transfected with almost no sign of cytotoxicity. Conclusions: Overall, SH-14 presents good potential as a novel transfection reagent to be used in the presence of serum.
AB - Background: Despite the use of currently optimized lipofection conditions, including transfection in serum-depleted media, the efficiency of gene transfer is low and high transfection rates often induce cytotoxicity. A lipid formulation with transfection efficiency not inhibited by serum would provide an advance towards in vivo applications. Methods: We explored the ability of the cationic lipid SH-14 to dimerize upon DNA and form lipoplexes, and potentially release nucleic acids in the intracellular reducing milieu. We investigated the critical micelle-forming concentration of SH-14 and its intrinsic toxicity, size and Zeta potential measurements, the in vitro cytotoxicity of SH-14/ plasmid DNA lipoplexes and their ability to transfect cells. Results: Among all the charge ratios (CR, +/-) tested, lipoplexes at CR 10 with a mean diameter of 295 nm and a surface charge of +20 mV, exhibited the best compromise between transfection efficiency and tolerability. SH-14 presented the same cytotoxicity level whether alone or complexed in lipoplexes. Lipofections carried out in serum-free medium shared a transfection efficiency, on average, of 40% and a cytotoxicity of 38%. An increase of 73% in transfection efficiency and 24% in cell viability were obtained, extending lipofection over 48 h in complete-medium. Moreover, when serum concentration was increased from 10% to 50%, a three-fold increase in plasmid dose led to more than 72% of cells being transfected with almost no sign of cytotoxicity. Conclusions: Overall, SH-14 presents good potential as a novel transfection reagent to be used in the presence of serum.
KW - Dimerizable function
KW - Gene delivery
KW - Lipoplex
KW - Nonviral vector
KW - Serum effect
KW - Transfection
UR - http://www.scopus.com/inward/record.url?scp=46649104752&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=46649104752&partnerID=8YFLogxK
U2 - 10.1002/jgm.1186
DO - 10.1002/jgm.1186
M3 - Article
C2 - 18338839
AN - SCOPUS:46649104752
VL - 10
SP - 637
EP - 645
JO - Journal of Gene Medicine
JF - Journal of Gene Medicine
SN - 1099-498X
IS - 6
ER -