Expression and activity of osteoblast-targeted Cre recombinase transgenes in murine skeletal tissues
Expression and activity of osteoblast-targeted Cre recombinase transgenes in murine skeletal tissues
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DOI：
10.1387/ijdb.041816fl
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发表时间：
2004-09-01
影响因子：
0.7
通讯作者：
Kream, BE
中科院分区：
文献类型：
--
作者：
Liu, F;Woitge, HW;Kream, BE
Cre/loxP重组系统可用于规避小鼠中广泛基因消除的许多限制。在这里，我们介绍了转基因小鼠的开发和表征，其中Cre重组酶已通过大鼠Col 1a 1启动子的2.3 kb（Col 2.3-Cre）和3.6 kb（Col 3.6-Cre）片段靶向成骨细胞谱系的细胞。Cre mRNA在成年Col 2.3-Cre和Col 3.6-Cre小鼠的颅骨和长骨以及Col 3.6-Cre小鼠的肌腱和皮肤中检测到。为了获得Cre介导的基因重排的时间和空间模式的历史标记，将Col-Cre小鼠与ROSA 26（R26 R）小鼠交配，其中Cre介导的floxed盒的切除导致LacZ表达。Col 2.3-Cre; R26 R和Col 3.6-Cre; R26 R后代、颅骨和长骨成骨细胞在胚胎第18天和出生后第5天显示出强烈的β-gal染色。P-gal染色的空间模式在骨和从Col 2.3-Cre; R26 R小鼠建立的骨髓基质培养物中更受限制。在携带Col 1a 1-GFP视觉报告基因的转基因骨中观察到表达空间模式的类似差异。我们的数据表明，Col 2.3-Cre和Col 3.6-Cre转基因小鼠可能用于体内条件基因靶向或用于获得成骨细胞群体用于体外培养，其中感兴趣的基因已被灭活。
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 beta-gal staining at embryonic day 18 and postnatal day 5. The spatial pattern of P-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 maybe 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.