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.

Indra AK, Warot X, Brocard J, Bornert JM, Xiao JH, Chambon P, Metzger D
Nucleic Acids Res. 1999 27 (22): 4324-7

PMID: 10536138 · PMCID: PMC148712 · DOI:10.1093/nar/27.22.4324

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).

MeSH Terms (15)

Animals Enzyme Induction Epidermis Estrogen Receptor Modulators Genes, Reporter Humans Integrases Keratinocytes Mice Mice, Transgenic Mutagenesis, Site-Directed Receptors, Estrogen Recombinant Fusion Proteins Tamoxifen Viral Proteins

Connections (1)

This publication is referenced by other Labnodes entities:

Links