Concept

Transfection

Publications associées (149)

Transfection using plasmid preparations

There is provided a method for plasmid DNA extraction to reproducibly yield relatively crude preparation which is suitable for transfection into mammalian cells. The inventors have determined that the high level of purification previously thought to be nec ...

2002

Lentiviral-mediated delivery of mutant huntingtin in the striatum of rats induces a selective neuropathology modulated by polyglutamine repeat size, huntingtin expression levels, and protein length

Patrick Aebischer, Diana Zala

A new strategy based on lentiviral-mediated delivery of mutant huntingtin (htt) was used to create a genetic model of Huntington's disease (HD) in rats and to assess the relative contribution of polyglutamine (CAG) repeat size, htt expression levels, and p ...

2002

Detection of promoter activity by flow cytometric analysis of GFP reporter expression

Joachim Lingner

Low efficiency of transfection is often the limiting factor for acquiring conclusive data in reporter assays. It is especially difficult to efficiently transfect and characterize promoters in primary human cells. To overcome this problem we have developed ...

2002

S-phase synchronized CHO cells show elevated transfection efficiency and expression using CaPi

Florian Maria Wurm, Martin Jordan, Frédéric Grosjean

Various methods exist to transfect mammalian cells in culture. It is generally accepted that individual methods have to be optimized for each of the cell lines or cell types used. Despitethe use of optimized protocols, significant day-to-day variationsin t ...

2002

Transfer of high copy number plasmid into mammalian cells by calcium phosphate transfection

Florian Maria Wurm, Martin Jordan

Using flow cytometry, single cell sorting, confocal microscopy and fluorescent plasmids, a thorough study of DNA uptake, DNA fate and DNA expression in mammalian cells transfected with the widely used calcium-phosphate precipitation method was executed. We ...

2001

Transient gene expression: recombinant protein production with suspension-adapted HEK293-EBNA cells

Florian Maria Wurm, Horst Pick, Klaus Friedrich

Transient gene expression (TGE) in mammalian cells at the reactor scale is becoming increasingly important for the rapid production of recombinant proteins. We improved a process for transient calcium phosphate-based transfection of HEK293-EBNA cells in a ...

2001

Analysis of the mouse Scnn1a promoter in cortical collecting duct cells and in transgenic mice

Friedrich Beermann, Sébastien Kohler

We have isolated and characterised the promoter of the mouse Scnn1a (alpha ENaC) gene. Using transient transfections of serial deletion mutants into Scnn1a-expressing cells, we demonstrate that 1.56 kb of 5' upstream sequence is required for cell-specific ...

2001

Development of stable cell lines for production or regulated expression using matrix attachment regions

Florian Maria Wurm

One of the major hurdles of isolating stable, inducible or constitutive high-level producer cell lines is the time-consuming selection procedure. Given the variation in the expression levels of the same construct in individual clones, hundreds of clones mu ...

2001

Inhibition of HIV-1 in cell culture by oligonucleotide-loaded nanoparticles

Didier Trono, Priscilla Turelli

PURPOSE: To investigate the potential use of polymeric nanoparticles for the delivery of antisense oligonucleotides in HIV-1-infected cell cultures. METHODS: Phosphorothioate oligonucleotides were encapsulated into poly (D,L-lactic acid) nanoparticles. Two ...

2001

Lentiviral-mediated gene transfer into haematopoietic stem cells

Didier Trono

OBJECTIVES: Lentiviral vectors can transduce nondividing cells. As most haematopoietic stem cells (HSCs) are nondividing in vivo, lentiviral vectors are promising viral vectors to transfer genes into HSCs. DESIGN AND SETTING: We have used HIV-1 based lenti ...

Wiley-Blackwell2001

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