Journal of Genetic Syndromes & Gene Therapy
Open Access
ISSN: ISSN: 2157-7412
ISSN: ISSN: 2157-7412
Seiichi Yamano
Scientific Tracks Abstracts: J Genet Syndr Gene Ther
To improve the efficiency of non-viral gene delivery, we combined the lipid FuGENE HD (FH) with the HIV Tat peptide sequence modified with histidine and cysteine residues (mTat), and evaluated whether mTat/FH could be used for gene delivery in vitro and in vivo . mTat/FH transfection was evaluated by luciferase expression plasmid in five cell types compared to five commercial reagents. Cytotoxicity, the zeta potentials, and size of mTat/FH and each component individually were determined, and transfection efficiency at different temperatures was examined. The endocytosis mechanism of mTat/FH/ DNA complexes was investigated. To examine the ability of mTat/FH in vivo , luciferase expression was measured using a real- time bioluminescence imaging system after intramuscular administration of mTat/FH/DNA. mTat/FH produced significant improvement in transfection efficiency of all cell lines with little cytotoxicity when compared to mTat alone, FH alone, or five commercial reagents. The zeta potential of mTat/FH/DNA was significantly higher compared to FH, mTat, or their DNA combination. The particle size of the FH/DNA complex was significantly reduced by addition of mTat. It was revealed that temperature-dependent and caveolae-mediated endocytosis of mTat/FH transfection was utilized. The results of in vivo study demonstrated the animals intramuscularly administered with mTat/FH/DNA had significantly higher and longer luciferase expression than those with mTat/DNA or FH/DNA. These findings demonstrated a combination of mTat with lipids improved transfection efficiency in several cell lines and it enabled efficient gene delivery in vivo. It makes mTat/FH a highly interesting non-viral gene vector for the future utilization, both in vitro and in vivo.
Seiichi Yamano is Assistant Professor of Prosthodontics at New York University College of Dentistry. Dr. Yamano first earned his dental degree from Nihon University and then a PhD in Medical Sciences from Tokyo Medical University in Japan. Then, he came to the US as a Research Fellow at the National Institutes of Health. Also, he received his DMD from the University of Pennsylvania, and a Prosthodontics certificate and MMSc in Oral Biology from Harvard University. Currently, his laboratory is focused on tissue engineering for oral and craniofacial regeneration using gene therapeutic technology, especially novel non-viral vectors.