Transcriptional and posttranslational regulation of Cre recombinase by RU486 as the basis for an enhanced inducible expression system

Stephanos Kyrkanides, Jen Nie H. Miller, William J. Bowers, Howard J. Federoff

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)790-795
Number of pages6
JournalMolecular Therapy
Volume8
Issue number5
DOIs
StatePublished - Nov 2003

Bibliographical note

Funding Information:
This work was supported in part by NIH Grants DE13860 (S.K.), DE00471 (S.K.), and MH57047 (H.J.F.) and an Orthodontic Faculty Development Fellowship Award from the American Association of Orthodontists Foundation to S.K. We also thank Dr. William H. Bowen for his constructive review of the manuscript.

Funding

This work was supported in part by NIH Grants DE13860 (S.K.), DE00471 (S.K.), and MH57047 (H.J.F.) and an Orthodontic Faculty Development Fellowship Award from the American Association of Orthodontists Foundation to S.K. We also thank Dr. William H. Bowen for his constructive review of the manuscript.

FundersFunder number
National Institutes of Health (NIH)DE00471, DE13860
National Institute of Mental HealthR01MH057047
American Association of Orthodontists Foundation

    Keywords

    • Cre
    • DNA
    • IoxP
    • Recombination
    • Site directed

    ASJC Scopus subject areas

    • Molecular Medicine
    • Molecular Biology
    • Genetics
    • Pharmacology
    • Drug Discovery

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