Int. J. Dev. Biol. 48: 645 - 653 (2004)

https://doi.org/10.1387/ijdb.041816fl

Vol 48, Issue 7

Expression and activity of osteoblast-targeted Cre recombinase transgenes in murine skeletal tissues

Original Article | Published: 1 September 2004

Fei Liu¹, Henning W. Woitge¹, Alen Braut², Mark S. Kronenberg³, Alexander C. Lichtler³, Mina Mina² and Barbara E. Kream*^,1,3

Departments of Medicine¹, Pediatric Dentistry² and Genetics and Developmental Biology³, University of Connecticut Health Center, Farmington, Connecticut, USA.

Abstract

The Cre/loxP recombination system can be used to circumvent many of the limitations of generalized gene ablation in mice. Here we present the development and characterization of transgenic mice in which Cre recombinase has been targeted to cells of the osteoblast lineage with 2.3 kb (Col 2.3-Cre) and 3.6 kb (Col 3.6-Cre) fragments of the rat Col1a1 promoter. Cre mRNA was detected in calvaria and long bone of adult Col 2.3-Cre and Col 3.6-Cre mice, as well as in tendon and skin of Col 3.6-Cre mice. To obtain a historical marking of the temporal and spatial pattern of Cre-mediated gene rearrangement, Col-Cre mice were bred with ROSA26 (R26R) mice in which Cre-mediated excision of a floxed cassette results in LacZ expression. In Col 2.3-Cre;R26R and Col 3.6-Cre;R26R progeny, calvarial and long bone osteoblasts showed intense β-gal staining at embryonic day 18 and postnatal day 5. The spatial pattern of β-gal staining was more restricted in bone and in bone marrow stromal cultures established from Col 2.3-Cre;R26R mice. Similar differences in the spatial patterns of expression were seen in transgenic bone carrying Col1a1-GFP visual reporters. Our data suggest that Col 2.3-Cre and Col 3.6-Cre transgenic mice may be useful for conditional gene targeting in vivo or for obtaining osteoblast populations for in vitro culture in which a gene of interest has been inactivated.

Keywords

Cre recombinase, Col1a1 promoter, osteoblast, transgenic mouse, Cre/loxP