The Lac repressor provides a reversible gene expression system in undifferentiated and differentiated embryonic stem cell

Control of mammalian gene promoters by the bacterial LacI repressor provides reversible regulation and dose-response levels of derepressed expression by the lactose analog isopropyl thiogalactose (IPTG). Here, we show that insertion of LacI-binding sites in the ubiquitous beta-actin promoter confers...

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Bibliographic Details
Published in:Cellular and molecular life sciences : CMLS 2005-07, Vol.62 (14), p.1605-1612
Main Authors: Caron, L, Prot, M, Rouleau, M, Rolando, M, Bost, F, Binétruy, B
Format: Article
Language:English
Subjects:
Actins - genetics
Animals
Binding sites
Cell Differentiation - genetics
Cell Line
Dose-Response Relationship, Drug
Gene Expression Regulation - drug effects
Gene Expression Regulation - genetics
Genetic Vectors - genetics
Isopropyl Thiogalactoside - pharmacology
Mice
Myocytes, Cardiac - cytology
Myocytes, Cardiac - metabolism
Neurons - cytology
Neurons - metabolism
Plasmids - genetics
Promoter Regions, Genetic - genetics
Proteins
Repressor Proteins - genetics
Stem cells
Stem Cells - cytology
Stem Cells - metabolism
Transgenes - genetics
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Summary:Control of mammalian gene promoters by the bacterial LacI repressor provides reversible regulation and dose-response levels of derepressed expression by the lactose analog isopropyl thiogalactose (IPTG). Here, we show that insertion of LacI-binding sites in the ubiquitous beta-actin promoter confers a strong and dose-dependent IPTG-regulatable expression of transiently transfected reporter genes in mouse embryonic stem (ES) cells expressing LacI. We established ES cell lines stably expressing reporter genes under inducible control and found a five- to tenfold IPTG induction of transgene expression. The kinetics of induction is rapid and stable, and can be rapidly reversed after IPTG removal. Importantly, this regulatable expression was maintained throughout the differentiation process of ES cells, and observed in individual differentiated cardiomyocyte-like cells and neuronal-like cells. This reversible system is the first to function from undifferentiated to individual well-differentiated ES cells, providing a very useful tool to understand molecular mechanisms underlying ES cell self-renewal, commitment and differentiation.
ISSN:1420-682X
1420-9071
DOI:10.1007/s00018-005-5123-2