Modified montmorillonite as vector for gene delivery.
Currently, gene delivery systems can be divided into two parts: viral or non-viral vectors. In general, viral vectors have a higher efficiency on gene delivery. However, they may sometimes provoke mutagenesis and carcinogenesis once re-activating in human body. Lots of non-viral vectors have been developed that tried to solve the problems happened on viral vectors. Unfortunately, most of non-viral vectors showed relatively lower transfection rate. The aim of this study is to develop a non-viral vector for gene delivery system. Montmorillonite (MMT) is one of clay minerals that consist of hydrated aluminum with Si-O tetrahedrons on the bottom of the layer and Al-O(OH)2 octahedrons on the top. The inter-layer space is about 12 A. The room is not enough to accommodate DNA for gene delivery. In the study, the cationic hexadecyltrimethylammonium (HDTMA) will be intercalated into the interlayer of MMT as a layer expander to expand the layer space for DNA accommodation. The optimal condition for the preparation of DNA-HDTMA-MMT is as follows: 1 mg of 1.5CEC HDTMA-MMT was prepared under pH value of 10.7 and with soaking time for 2 h. The DNA molecules can be protected from nuclease degradation, which can be proven by the electrophoresis analysis. DNA was successfully transfected into the nucleus of human dermal fibroblast and expressed enhanced green fluorescent protein (EGFP) gene with green fluorescence emission. The HDTMA-MMT has a great potential as a vector for gene delivery in the future.
Lin
Feng-Huei
FH
Institute of Biomedical Engineering, National Taiwan University, and National Taiwan University Veternary Hospital, Taipei, Taiwan.
Chen
Chia-Hao
CH
Cheng
Winston T K
WT
Kuo
Tzang-Fu
TF
D000535:Aluminum / Q000737:chemistry; D001546:Bentonite / Q000494:pharmacology*; D002412:Cations; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry; D003851:Deoxyribonucleases / Q000737:chemistry / Q000378:metabolism; D004586:Electrophoresis; D004705:Endocytosis; D005347:Fibroblasts / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D012825:Silicon / Q000737:chemistry; D013696:Temperature; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection
D002412:Cations; D013778:Tetrazolium Salts; D013844:Thiazoles; D001546:Bentonite; D049452:Green Fluorescent Proteins; D004247:DNA; D000535:Aluminum; D003851:Deoxyribonucleases; C022616:thiazolyl blue; D012825:Silicon