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 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.
Caron
L
L
INSERM U568, Faculté de Médecine, Université de Nice Sophia-Antipolis, Avenue de Valombrose, 06107, Nice Cedex, France.
Prot
M
M
Rouleau
M
M
Rolando
M
M
Bost
F
F
Binétruy
B
B
D000199:Actins / Q000235:genetics; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics*; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D019076:Transgenes / Q000235:genetics
D000199:Actins; D012097:Repressor Proteins; D007544:Isopropyl Thiogalactoside