S4₁₃-PV cell penetrating peptide and cationic liposomes act synergistically to mediate intracellular delivery of plasmid DNA

TY - JOUR

T1 - S413-PV cell penetrating peptide and cationic liposomes act synergistically to mediate intracellular delivery of plasmid DNA

AU - Trabulo, Sara

AU - Mano, Miguel

AU - Faneca, Henrique

AU - Cardoso, Ana Luísa

AU - Duarte, Sónia

AU - Henriques, Ana

AU - Paiva, Artur

AU - Gomes, Paula

AU - Simões, Sérgio

AU - Pedroso de Lima, Maria C.

PY - 2008

Y1 - 2008

N2 - Background: Cell penetrating peptides have been successfully used to mediate the intracellular delivery of a wide variety of molecules of pharmacological interest. The main aim of the present work was to evaluate the potential of the S413-PV cell penetrating peptide to mediate the intracellular delivery of plasmid DNA, aiming at its use in gene therapy applications. The S413-PV cell penetrating peptide is a chimeric peptide that results from the combination of a cell penetrating sequence derived from the Dermaseptin S4 peptide with the nuclear localization signal present in the Simian Virus 40 (SV40) large T antigen. Methods: S413-PV cell penetrating peptide and cationic liposomes composed of 1,2-dioleoyl-3-trimethylammonium-propane: 1,2-dioleoyl-snglycero-3-phosphoethanolamine were complexed with pDNA at different charge ratios. Complexation of pDNA was assessed by gel electrophoresis. Luciferase assay, fluorescence microscopy and fluorescence-activated cell sorting analysis were used to evaluate reporter gene delivery to TSA and HeLa cells. Cytotoxicity of the pDNA complexes was assessed by Alamar blue assay. Results: Complexes obtained through electrostatic association of the S413-PV cell penetrating peptide with plasmid DNA are able to very efficiently mediate transfection, particularly at high peptide/ DNA charge ratios. Additionally, our results clearly demonstrate that, both in HeLa and TSA cells, ternary complexes, resulting from association of cationic liposomes to peptide/DNA complexes, are significantly more efficient in mediating transfection than the corresponding peptide/DNA or cationic liposome/DNA complexes. Conclusions: Overall, our data highlight the potential of cell penetrating peptides for the development of improved nonviral gene delivery systems.

AB - Background: Cell penetrating peptides have been successfully used to mediate the intracellular delivery of a wide variety of molecules of pharmacological interest. The main aim of the present work was to evaluate the potential of the S413-PV cell penetrating peptide to mediate the intracellular delivery of plasmid DNA, aiming at its use in gene therapy applications. The S413-PV cell penetrating peptide is a chimeric peptide that results from the combination of a cell penetrating sequence derived from the Dermaseptin S4 peptide with the nuclear localization signal present in the Simian Virus 40 (SV40) large T antigen. Methods: S413-PV cell penetrating peptide and cationic liposomes composed of 1,2-dioleoyl-3-trimethylammonium-propane: 1,2-dioleoyl-snglycero-3-phosphoethanolamine were complexed with pDNA at different charge ratios. Complexation of pDNA was assessed by gel electrophoresis. Luciferase assay, fluorescence microscopy and fluorescence-activated cell sorting analysis were used to evaluate reporter gene delivery to TSA and HeLa cells. Cytotoxicity of the pDNA complexes was assessed by Alamar blue assay. Results: Complexes obtained through electrostatic association of the S413-PV cell penetrating peptide with plasmid DNA are able to very efficiently mediate transfection, particularly at high peptide/ DNA charge ratios. Additionally, our results clearly demonstrate that, both in HeLa and TSA cells, ternary complexes, resulting from association of cationic liposomes to peptide/DNA complexes, are significantly more efficient in mediating transfection than the corresponding peptide/DNA or cationic liposome/DNA complexes. Conclusions: Overall, our data highlight the potential of cell penetrating peptides for the development of improved nonviral gene delivery systems.

KW - Cationic liposomes

KW - Cell penetrating peptide

KW - Gene technology

KW - Nonviral vectors

KW - Nuclear localization signal

KW - Plasmid DNA

UR - http://www.scopus.com/inward/record.url?scp=58149190435&partnerID=8YFLogxK

U2 - 10.1002/jgm.1247

DO - 10.1002/jgm.1247

M3 - Article

C2 - 18729238

AN - SCOPUS:58149190435

SN - 1099-498X

VL - 10

SP - 1210

EP - 1222

JO - JOURNAL OF GENE MEDICINE

JF - JOURNAL OF GENE MEDICINE

IS - 11

ER -