Optimization of Procedure for Efficient Gene Transfer into Porcine Somatic Cells with Lipofection

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Optimization of Procedure for Efficient Gene Transfer into Porcine Somatic Cells with Lipofection

The objective of this study was to establish conditions for transfection of a foreign gene into somatic cells using cationic lipid reagents and to evaluate the effects of transfection on in vitro development of somatic cell nuclear transfer (SCNT) embryos. Green fluorescent protein (GFP) gene was used as a foreign gene and a non-transfected somatic cell was utilized as a control karyoplast. Mon...

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Gene Transfer into Sub- cultured Endometrial Cells Using Lipofection

Lipofection using the Lipofectin® reagent was optimized to transiently transfect subcultured guinea pig endometrial stromal cells with a β-galactosidase gene driven by a simian virus 40 promoter. Efficient transfection was obtained in the following conditions: a value of six for the ratio of lipofectin to DNA, a low cellular density (105 cells per 35-mm well) at the time of subculture (48 h bef...

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Gene transfer into subcultured endometrial cells using lipofection.

Lipofection using the Lipofectin reagent was optimized to transiently transfect subcultured guinea pig endometrial stromal cells with a beta-galactosidase gene driven by a simian virus 40 promoter. Efficient transfection was obtained in the following conditions: a value of six for the ratio of lipofectin to DNA, a low cellular density (10(5) cells per 35-mm well) at the time of subculture (48 h...

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Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure.

A DNA-transfection protocol has been developed that makes use of a synthetic cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTMA). Small unilamellar liposomes containing DOTMA interact spontaneously with DNA to form lipid-DNA complexes with 100% entrapment of the DNA, DOTMA facilitates fusion of the complex with the plasma membrane of tissue culture cells, resul...

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Dynamic Reorganization of Nucleosome Positioning in Somatic Cells after Transfer into Porcine Enucleated Oocytes

The nucleosome, the fundamental structural unit of chromatin, is a critical regulator of gene expression. The mechanisms governing changes to nucleosome occupancy and positioning during somatic cell reprogramming remain poorly understood. We established a method for generating genome-wide nucleosome maps of porcine embryonic fibroblasts (PEF), reconstructed 1-cell embryos generated by somatic c...

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ژورنال

عنوان ژورنال: Asian-Australasian Journal of Animal Sciences

سال: 2008

ISSN: 1011-2367,1976-5517

DOI: 10.5713/ajas.2008.70400