Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10904837 | Experimental Cell Research | 2005 | 16 Pages |
Abstract
We demonstrate a cell extract-based, genome-wide and heritable reprogramming of gene expression in vitro. Kidney epithelial 293T cells have previously been shown to take on T cell properties following a brief treatment with an extract of Jurkat T cells. We show here that 293T cells exposed for 1 h to a Jurkat cell extract undergo genome-wide, target cell-type-specific and long-lasting transcriptional changes. Microarray analyses indicate that on any given week after extract treatment, â¼2500 genes are upregulated >3-fold, of which â¼900 are also expressed in Jurkat cells. Concomitantly, â¼1500 genes are downregulated or repressed, of which â¼500 are also downregulated in Jurkat cells. Gene expression changes persist for over 30 passages (â¼80 population doublings) in culture. Target cell-type specificity of these changes is shown by the lack of activation or repression of Jurkat-specific genes by extracts of 293T cells or carcinoma cells. Quantitative RT-PCR analysis confirms the long-term transcriptional activation of genes involved in key T cell functions. Additionally, growth of cells in suspended aggregates, expression of CD3 and CD28 T cell surface markers, and interleukin-2 secretion by 293T cells treated with extract of adult peripheral blood T cells illustrate a functional nuclear reprogramming. Therefore, target cell-type-specific and heritable changes in gene expression, and alterations in cell function, can be promoted by extracts derived from transformed cells as well as from adult primary cells.
Keywords
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Authors
Anne-Mari HÃ¥kelien, Kristine G. Gaustad, Christel K. Taranger, Bjørn S. SkÃ¥lhegg, Thomas Küntziger, Philippe Collas,