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In vitro and in vivo effects of a multimerized αs1‐casein enhancer on whey acidic protein gene promoter activity
Author(s) -
Pantano Thais,
RivalGervier Sylvie,
Prince Sonia,
Bourhis Celeste MenckLe,
Maeder Caroline,
Viglietta Céline,
Houdebine LouisMarie,
Jolivet Geneviève
Publication year - 2003
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.10302
Subject(s) - enhancer , luciferase , biology , microbiology and biotechnology , reporter gene , transgene , gene silencing , gene , genetically modified mouse , transfection , gene expression , biochemistry
Experimental data obtained in previous works have led to postulate that enhancers increase the frequency of action of a linked promoter in a given cell and may have some insulating effects. The multimerized rabbit αs1‐casein gene enhancer, the 6i multimer, was added upstream of the rabbit whey acidic protein gene (WAP) promoter (−6,300; +28 bp) fused to the firefly luciferase ( luc ) gene (6i WAP‐luc construct). The 6i multimer increased reporter gene expression in mouse mammary HC11 cells. In transgenic mice, a very weak but significant increase was also observed. More noticeable, no silent lines were found when the 6i multimer was associated to the WAP‐luc construct. This reflects the fact that the 6i multimer tends to prevent the silencing of the WAP‐luc construct. After addition of the 5′HS4 insulator region from the chicken β‐globin locus upstream of the 6i multimer, similar luciferase levels were measured in 6i WAP‐luc and 5′HS4 WAP‐luc transgenic mice. Our present data and previous ones, which show that the 6i multimer has no insulating activity on a TK gene promoter construct indicate that the insulating activity of the 6i multimer is construct‐dependent and not amplified by the 5′HS4 insulator. Mol. Reprod. Dev. 65: 262–268, 2003. © 2003 Wiley‐Liss, Inc.