Transcriptional and posttranslational regulation of Cre recombinase by RU486 as the basis for an enhanced inducible expression system.
Genetic studies often require the employment of an inducible expression system, whereby the expression of a particular gene can be regulated by the exogenous administration of an inert ligand. Cre/loxP-based systems have been previously described as the basis for inducible expression systems by exerting site-specific DNA recombination. In our effort to enhance the properties of the RU486-responsive CrePr1 construct, we have developed the dual GLVP/CrePr system, in which RU486 confers activity control at both the transcriptional and the posttranslational level of CrePr1. This was achieved by placing CrePr1 transcriptional regulation under the control of the RU486-sensitive chimeric regulator GLVP. Stable cell lines harboring the dual GLVP/CrePr as well as the single CrePr1 system were developed. Our results indicate that the dually regulated system is highly inducible by RU486 while maintaining minimal basal activity ("leakage"), characteristics that can be employed in the development of transgenic mice in which genetic pathways can be turned on or turned off after exogenous administration of RU486 at physiologically inert doses.
Kyrkanides
Stephanos
S
Department of Dentistry, Rochester, New York 14620, USA. Stephanos_Kyrkanides@urmc.rochester.edu
Miller
Jen nie H
Jn
Bowers
William J
WJ
Federoff
Howard J
HJ
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006727:Hormone Antagonists / Q000627:therapeutic use; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D015735:Mifepristone / Q000627:therapeutic use*; D008957:Models, Genetic; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011995:Recombination, Genetic; D013997:Time Factors; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D006727:Hormone Antagonists; D014764:Viral Proteins; D015735:Mifepristone; C045073:Cre recombinase; D019426:Integrases