Non-toxic phototriggered gene transfection by PAMAM-porphyrin conjugates.
Development of controllable and non-toxic gene transfection systems is a core issue in gene therapy. Photochemical internalization, an innovative strategy in cytosolic release, provides us with an opportunity to develop a light-inducible gene delivery system. In this study, a novel photochemical internalization (PCI)-mediated gene delivery system was synthesized by surface modification of polyamidoamine (PAMAM) dendrimers via 5,10,15-tri(4-acetamidophenyl)-20-mono(4-carboxyl-phenyl)porphyrin (TAMCPP) conjugated to the generation 4 PAMAM dendrimer (G4). This water-soluble PAMAM-TAMCPP conjugate was characterized for cell viability, phototoxicity, DNA complexation, and in vitro transfection activity. The results show that TAMCPP conjugation did not increase the cytotoxicity of the PAMAM dendrimer below 20 microM, but significantly induced cell death after suitable irradiation. Under almost non-toxic G4-TAMCPP-mediated PCI treatment, the expression of green fluorescent protein determined by flow cytometry could be markedly enhanced in HeLa cells. Therefore, the G4-TAMCPP conjugate had an inducible and effective gene transfection activity, and showed considerable potential as a bimodal biomaterial for PCI-mediated gene therapy.
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, No. 1, Section 1, Jen-Ai Road, Taipei 100, Taiwan.
Ming-Jium
MJ
Shieh
Cheng-Liang
CL
Peng
Pei-Jen
PJ
Lou
Chieh-Hua
CH
Chiu
Tsiao-Yu
TY
Tsai
Chia-Yen
CY
Hsu
Chen-Yu
CY
Yeh
Ping-Shan
PS
Lai
D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism; D050091:Dendrimers / Q000009:adverse effects / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D005434:Flow Cytometry; D005470:Fluorometry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism; D015394:Molecular Structure; D009757:Nylons / Q000009:adverse effects / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D000442:Octanols / Q000737:chemistry; D010778:Photochemotherapy; D011166:Porphyrins / Q000009:adverse effects / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D012995:Solubility; D013778:Tetrazolium Salts / Q000378:metabolism; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D014867:Water / Q000737:chemistry
D050091:Dendrimers; D007202:Indicators and Reagents; D009757:Nylons; D000442:Octanols; C520550:PAMAM-G4; D011166:Porphyrins; D013778:Tetrazolium Salts; D014867:Water; D049452:Green Fluorescent Proteins; D004247:DNA