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TURAN, KADİR

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KADİR

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2006 - 200912010 - 20161
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    Chitosan-DNA nanoparticles: The effect of cell type and hydrolysis of chitosan on in vitro DNA transfection
    (TAYLOR & FRANCIS INC, 2006) TURAN, KADİR; Turan, Kadir; Nagata, Kyosuke
    Commercial chitosan (Ch) with low (LMWCh) and medium molecular weight (MMWCh) were hydrolyzed in diluted hydrochloric acid by heating at different temperatures. The viscosity average molecular weight of Chs was gradually decreased from 450 to 14 kDa as a function of temperature. Ch fractions were used for formation of Ch-DNA nanoparticles and tested for the ability to introduce DNA into HEK293, Swiss3T3, HeLa, and MDCK cells in vitro. The average diameter of nanoparticles was 200-220 nm. The surface charge of nanoparticles varied depending on the Ch/DNA ratio. The cell lines different response to DNA transection with Ch fractions depended on molecular weight. HEK293 cells were efficiently transfected by nanoparticles prepared with Chs having a wide range of molecular weight (similar to 14-195 kDa). Swiss3T3 cells were efficiently transfected by Ch polymers with about <17 kDa. In contrast, HeLa and MDCK cells were highly resistant to DNA transfection with Ch polymers. These results strongly suggest that Ch polymers may be widely used for DNA trasnfection of the mammalian cells under optimized conditions.
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    The potential of archaeosomes as carriers of pDNA into mammalian cells
    (TAYLOR & FRANCIS LTD, 2016) OGAN, AYŞE; Attar, Azade; Ogan, Ayse; Yucel, Sevil; Turan, Kadir
    This paper describes the formulation of archaeosomes and the evaluation of their abilities to facilitate in vitro DNA delivery. Lipids of the H.hispanica 2TK2 strain were used in archaeosome formation, which is formulated by mixing H.hispanica 2TK2 lipids with plasmid DNA encoding green fluorescent protein (GFP) or beta-galactosidase (beta-gal). Archaeosome/pDNA formation and unbound DNA were monitored by agarose gel electrophoresis. The archaeosome formulations were visualized by AFM and TEM. The zeta potential analysis showed the archaeosomes to be electronegative. The composition of archaeosomes and the DNA dose for transient transfection into HEK293 cells were optimized, and the relationship between the structure and activity of archaeosomes in DNA delivery was investigated. By themselves, archaeosomes showed low efficiency for DNA delivery, due to their anionic nature. By formulating archaeosomes with a helper molecule, such as DOTAP, CaCl2, or LiCl, the capability of archaeosomes for gene transfection is significantly enhanced. The transfection profiles of efficient archaeosomes are proved to have a long shelf-life when maintained at room temperature. Thus, the archaeal lipids have the potential to be used as transfection reagents in vitro.