Improved reporter strain for monitoring Cre recombinase-mediated DNA excisions in mice
Improved reporter strain for monitoring Cre recombinase-mediated DNA excisions in mice
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DOI：
10.1073/pnas.96.9.5037
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发表时间：
1999-04-27
影响因子：
11.1
通讯作者：
Orkin, SH
中科院分区：
文献类型：
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作者：
Mao, XH;Fujiwara, Y;Orkin, SH
有效利用条件性Cre重组酶 - loxP基因修饰需要具有明确表达模式的Cre表达小鼠品系。为了评估Cre表达小鼠的体内功能，我们构建了一种改进的报告基因品系，用于监测Cre介导的切除作用。通过基因打靶对β - 半乳糖苷酶 - 新霉素磷酸转移酶融合基因（βgeo）捕获的ROSA26基因座进行了修饰，使得只有在Cre介导切除loxP侧翼的DNA序列后βgeo才会表达。从切除的ROSA26等位基因产生的βgeo在胚胎和成年小鼠中普遍表达。通过将报告基因品系与Cre表达转基因小鼠交配，我们表明在早期胚胎发生过程中以及在特定的造血谱系（T淋巴细胞）中，loxP侧翼的ROSA26等位基因可被Cre作用。这种改进的报告基因品系应有助于在各种实验环境中监测体内Cre介导的切除事件。
Effective use of conditional Cre recombinase-loxP gene modification requires Cre-expressing mouse strains with defined patterns of expression. To assess the in vivo functionality of Cre expressing mice, we have engineered an improved reporter strain for monitoring Cre-mediated excisions. The beta-galactosidase-neomycin phosphotransferase fusion gene (beta geo)-trapped ROSA26 locus was modified by gene targeting such that beta geo is expressed only after Cre-mediated excision of loxP-flanked DNA sequences. beta geo from the excised ROSA26 allele is expressed ubiquitously in embryos and adult mice. By mating the reporter strain with Cre-expressing transgenic mice, we have shown that the loxP-flanked ROSA26 allele is accessible to Cre during early embryogenesis, as well as in a specific hematopoietic lineage (T lymphocytes). This improved reporter strain should facilitate monitoring in vivo Cre-mediated excision events in a variety of experimental contexts.