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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Improved reporter strain for monitoring Cre recombinase- mediated DNA excisions in mice
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Improved reporter strain for monitoring Cre recombinase- mediated DNA excisions in mice

机译：改良的报告株，用于监测小鼠中Cre重组酶介导的DNA切除

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摘要

Effective use of conditional Cre recombi- nase-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 axcisions. 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 em- bryos 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 Iymphocytes). This improved reporter strain should facil- itate monitoring in vivo Cre-mediated excision events in a variety of cxperimental contexts.
机译：有效使用条件Cre重组酶loxP基因修饰需要表达Cre的小鼠品系具有明确的表达模式。为了评估表达Cre的小鼠的体内功能，我们设计了一种改良的报告株，用于监测Cre介导的脱落。捕获＃beta＃-半乳糖苷酶-新霉素磷酸转移酶融合基因（＃beta＃geo）的ROSA26基因座，通过基因靶向修饰，使得仅在Cre介导的loxP侧翼DNA序列切除后才表达＃beta＃geo。切除的ROSA26等位基因的＃beta＃geo在胚胎和成年小鼠中普遍表达。通过使报告菌株与表达Cre的转基因小鼠交配，我们已经表明，loxP侧翼的ROSA26等位基因在早期胚胎发生期间以及在特定的造血谱系（T淋巴细胞）中均可被Cre接近。这种改进的报告株应有助于在各种实验环境中监测体内Cre介导的切除事件。

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