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

被引:8
作者
Caron, L
Prot, M
Rouleau, M
Rolando, M
Bost, F
Binétruy, B
机构
[1] Univ Nice, Fac Med, INSERM, U568, F-06107 Nice, France
[2] Univ Nice, Fac Med, INSERM, U634, F-06107 Nice, France
关键词
mouse embryonic stem cell; LacI/IPTG reversible expression vector; ubiquitous gene promoter; neuron; cardiomyocyte;
D O I
10.1007/s00018-005-5123-2
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 P-actin promoter confers a strong and dose-dependent IPTG-regulatable expression of transiently transfected reporter genes in mouse embryonic stem (ES) cells expressing Lacl. 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.
引用
收藏
页码:1605 / 1612
页数:8
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