Temporally-controlled site-specific mutagenesis in the basal layer of the epidermis: comparison of the recombinase activity of the tamoxifen-inducible Cre-ER(T) and Cre-ER(T2) recombinases

Conditional DNA excision between two LoxP sites can be achieved in the mouse using Cre-ER(T), a fusion protein between a mutated ligand binding domain of the human estrogen receptor (ER) and the Cre recombinase, the activity of which can be induced by 4-hydroxy-tamoxifen (OHT), but not natural ER li...

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Bibliographic Details
Published in:Nucleic acids research 1999-11, Vol.27 (22), p.4324-4327
Main Authors: Indra, A K, Warot, X, Brocard, J, Bornert, J M, Xiao, J H, Chambon, P, Metzger, D
Format: Article
Language:English
Subjects:
Animals
Biochemistry, Molecular Biology
Enzyme Induction
Epidermis - drug effects
Epidermis - enzymology
Estrogen Receptor Modulators - pharmacology
Genes, Reporter
Humans
Integrases - biosynthesis
Integrases - genetics
Integrases - metabolism
Keratinocytes - drug effects
Keratinocytes - physiology
Life Sciences
Mice
Mice, Transgenic
Mutagenesis, Site-Directed
Receptors, Estrogen - antagonists & inhibitors
Receptors, Estrogen - genetics
Receptors, Estrogen - metabolism
Recombinant Fusion Proteins - drug effects
Recombinant Fusion Proteins - metabolism
Tamoxifen - analogs & derivatives
Tamoxifen - pharmacology
Viral Proteins
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Summary:Conditional DNA excision between two LoxP sites can be achieved in the mouse using Cre-ER(T), a fusion protein between a mutated ligand binding domain of the human estrogen receptor (ER) and the Cre recombinase, the activity of which can be induced by 4-hydroxy-tamoxifen (OHT), but not natural ER ligands. We have recently characterized a new ligand-dependent recombinase, Cre-ER(T2), which was approximately 4-fold more efficiently induced by OHT than Cre-ER(T) in cultured cells. In order to compare the in vivo efficiency of these two ligand-inducible recombinases to generate temporally-controlled somatic mutations, we have engineered transgenic mice expressing a LoxP-flanked (floxed) transgene reporter and either Cre-ER(T) or Cre-ER(T2) under the control of the bovine keratin 5 promoter that is specifically active in the epidermis basal cell layer. No background recombinase activity could be detected, while recombination was induced in basal keratinocytes upon OHT administration. Interestingly, a dose-response study showed that Cre-ER(T2) was approximately 10-fold more sensitive to OHT induction than Cre-ER(T).
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/27.22.4324