胶原蛋白1α1启动子驱动转基因小鼠成骨细胞中Cre重组酶的表达。

Collagen1alpha1 promoter drives the expression of Cre recombinase in osteoblasts of transgenic mice.

作者信息

Zha Lagabaiyila, Hou Ning, Wang Jian, Yang Guan, Gao Yuanrong, Chen Lin, Yang Xiao

机构信息

State Key Laboratory of Proteomics, Genetic Laboratory of Development and Diseases, Institute of Biotechnology, Beijing, China.

出版信息

J Genet Genomics. 2008 Sep;35(9):525-30. doi: 10.1016/S1673-8527(08)60072-7.

DOI:10.1016/S1673-8527(08)60072-7

PMID:18804071

Abstract

Osteoblasts participate in bone formation, bone mineralization, osteoclast differentiation and many pathological processes. To study the function of genes in osteoblasts using Cre-LoxP system, we generated a mouse line expressing the Cre recombinase under the control of the rat Collagen1alpha1 (Col1alpha1) promoter (Col1alpha1-Cre). Two founders were identified by genomic PCR from 16 offsprings, and the integration efficiency is 12.5%. In order to determine the tissue distribution and the activity of Cre recombinase in the transgenic mice, the Col1alpha1-Cre transgenic mice were bred with the ROSA26 reporter strain and a mouse strain that carries Smad4 conditional alleles (Smad4(Co/Co)). Multiple tissue PCR of Col1alpha1-Cre;Smad4(Co/+)mice revealed the restricted Cre activity in bone tissues containing osteoblasts and tendon. LacZ staining in the Col1alpha1-Cre;ROSA26 double transgenic mice revealed that the Cre recombinase began to express in the osteoblasts of calvaria at E14.5. Cre activity was observed in the osteoblasts and osteocytes of P10 double transgenic mice. All these data indicated that the Col1alpha1-Cre transgenic mice could serve as a valuable tool for osteoblast lineage analysis and conditional gene knockout in osteoblasts.

摘要

成骨细胞参与骨形成、骨矿化、破骨细胞分化以及许多病理过程。为了利用Cre-LoxP系统研究成骨细胞中基因的功能，我们构建了一种在大鼠胶原蛋白1α1（Col1α1）启动子控制下表达Cre重组酶的小鼠品系（Col1α1-Cre）。通过基因组PCR从16只后代中鉴定出两个奠基者，整合效率为12.5%。为了确定转基因小鼠中Cre重组酶的组织分布和活性，将Col1α1-Cre转基因小鼠与ROSA26报告菌株以及携带Smad4条件等位基因的小鼠品系（Smad4(Co/Co)）进行杂交。对Col1α1-Cre;Smad4(Co/+)小鼠的多个组织进行PCR分析，结果显示在含有成骨细胞和肌腱的骨组织中Cre活性受到限制。对Col1α1-Cre;ROSA26双转基因小鼠进行LacZ染色，结果表明在胚胎第14.5天，Cre重组酶开始在颅盖骨的成骨细胞中表达。在出生后第10天的双转基因小鼠的成骨细胞和骨细胞中观察到Cre活性。所有这些数据表明，Col1α1-Cre转基因小鼠可作为成骨细胞谱系分析和成骨细胞中条件性基因敲除的有价值工具。