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

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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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Liposome plasmid DNA complexes (lipoplexes) are often inefficient in mediating gene transfer and expression because of DNA degradation in lysosomal vesicles. Because herpes simplex virus (HSV) enters cells by fusion of the virus envelope with the plasma membranes, thereby overriding the endosomal pathway, HSV/lipoplex mixtures could be useful for improving gene transfer particularly when the mi...

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A Simple, Highly Efficient Method for Heterologous Expression in Mammalian Primary Neurons Using Cationic Lipid-mediated mRNA Transfection

Expression of heterologous proteins in adult mammalian neurons is a valuable technique for the study of neuronal function. The post-mitotic nature of mature neurons prevents effective DNA transfection using simple, cationic lipid-based methods. Adequate heterologous protein expression is often only achievable using complex techniques that, in many cases, are associated with substantial toxicity...

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Highly Efficient Reverse siRNA Transfection of Human Epithelial Lung Carcinoma A549 Cells Using siLentFect Lipid Reagent

Methods Transfection A549 cells (American Type Culture Collection, ATCC #CCL-185) were grown in F-12K nutrient mixture, Kaighn’s modification (Invitrogen Corporation) with 10% fetal bovine serum. Transfection complexes were prepared in Opti-MEM I reduced-serum medium (Invitrogen). Human GAPDH siRNA and nonsilencing siRNA were used (siLentMer starter kit for human GAPDH, Bio-Rad Laboratories, In...

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ژورنال

عنوان ژورنال: Proceedings of the National Academy of Sciences

سال: 1987

ISSN: 0027-8424,1091-6490

DOI: 10.1073/pnas.84.21.7413