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Title: Long-term in vitro, cell-type-specific genome-wide reprogramming of gene expression

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, {approx}2500 genes are upregulated >3-fold, of which {approx}900 are also expressed in Jurkat cells. Concomitantly, {approx}1500 genes are downregulated or repressed, of which {approx}500 are also downregulated in Jurkat cells. Gene expression changes persist for over 30 passages ({approx}80 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 changesmore » in gene expression, and alterations in cell function, can be promoted by extracts derived from transformed cells as well as from adult primary cells.« less

Authors:
 [1];  [1];  [1];  [2];  [1];  [1]
  1. Institute of Basic Medical Sciences, Department of Biochemistry, University of Oslo, PO Box 1112 Blindern, 0317 Oslo (Norway)
  2. Institute of Basic Medical Sciences, Department of Nutrition Research, University of Oslo, PO Box 1112 Blindern, 0317 Oslo (Norway)
Publication Date:
OSTI Identifier:
20717649
Resource Type:
Journal Article
Journal Name:
Experimental Cell Research
Additional Journal Information:
Journal Volume: 309; Journal Issue: 1; Other Information: DOI: 10.1016/j.yexcr.2005.06.001; PII: S0014-4827(05)00270-3; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0014-4827
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; BLOOD; CARCINOMAS; GENES; IN VITRO; KIDNEYS; POLYMERASE CHAIN REACTION; SECRETION; TRANSCRIPTION

Citation Formats

Hakelien, Anne-Mari, Gaustad, Kristine G, Taranger, Christel K, Skalhegg, Bjorn S, Kuentziger, Thomas, and Collas, Philippe. Long-term in vitro, cell-type-specific genome-wide reprogramming of gene expression. United States: N. p., 2005. Web. doi:10.1016/j.yexcr.2005.06.001.
Hakelien, Anne-Mari, Gaustad, Kristine G, Taranger, Christel K, Skalhegg, Bjorn S, Kuentziger, Thomas, & Collas, Philippe. Long-term in vitro, cell-type-specific genome-wide reprogramming of gene expression. United States. https://doi.org/10.1016/j.yexcr.2005.06.001
Hakelien, Anne-Mari, Gaustad, Kristine G, Taranger, Christel K, Skalhegg, Bjorn S, Kuentziger, Thomas, and Collas, Philippe. 2005. "Long-term in vitro, cell-type-specific genome-wide reprogramming of gene expression". United States. https://doi.org/10.1016/j.yexcr.2005.06.001.
@article{osti_20717649,
title = {Long-term in vitro, cell-type-specific genome-wide reprogramming of gene expression},
author = {Hakelien, Anne-Mari and Gaustad, Kristine G and Taranger, Christel K and Skalhegg, Bjorn S and Kuentziger, Thomas and Collas, Philippe},
abstractNote = {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, {approx}2500 genes are upregulated >3-fold, of which {approx}900 are also expressed in Jurkat cells. Concomitantly, {approx}1500 genes are downregulated or repressed, of which {approx}500 are also downregulated in Jurkat cells. Gene expression changes persist for over 30 passages ({approx}80 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.},
doi = {10.1016/j.yexcr.2005.06.001},
url = {https://www.osti.gov/biblio/20717649}, journal = {Experimental Cell Research},
issn = {0014-4827},
number = 1,
volume = 309,
place = {United States},
year = {Sat Sep 10 00:00:00 EDT 2005},
month = {Sat Sep 10 00:00:00 EDT 2005}
}