Detection of targeted GFP-Hox gene fusions during mouse embryogenesis
Detection of targeted GFP-Hox gene fusions during mouse embryogenesis
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DOI：
10.1073/pnas.95.22.13042
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发表时间：
1998-10-27
影响因子：
11.1
通讯作者：
Capecchi, MR
中科院分区：
文献类型：
--
作者：
Godwin, AR;Stadler, HS;Capecchi, MR
使用重要细胞标记来研究小鼠胚胎发生的能力将为实验研究开辟新的途径。最近，使用含有多个编码绿色荧光蛋白（GFP）的水母基因的转基因小鼠已经开始实现这一潜力。在这里，我们表明，单拷贝、靶向 GFP 与两种不同的小鼠 Hox 基因 Hoxa1 和 Hoxc13 框内融合产生的荧光信号，可以通过使用共聚焦显微镜轻松检测到。由于Hoxa1在小鼠胚胎发生早期表达，而Hoxc13在小鼠胚胎发生晚期表达，本研究表明单拷贝GFP基因融合可用于大部分小鼠胚胎发生。此前，靶向 lacZ 基因融合对于分析小鼠突变体非常有用。 GFP 基因融合体的使用通过提供重要标记的额外效用，扩展了靶向 lacZ 基因融合体的优势。我们对 Hoxc13(GFPneo) 胚胎的分析揭示了从 Hoxc13(lacZneo) 胚胎分析中预期的每个位点中的 GFP 表达。同样，在 RNA 原位分析预测的所有位点中均检测到 Hoxa1(GFPneo) 表达。 Hoxa1 (GFPneo) 胚胎前肠袋中的 GFP 表达表明 Hoxa1 在前肠介导的心源性中胚层分化中发挥作用。
The ability to use a vital cell marker to study mouse embryogenesis will open new avenues of experimental research. Recently, the use of transgenic mice, containing multiple copies of the jellyfish gene encoding the green fluorescent protein (GFP), has begun to realize this potential. Here, we show that the fluorescent signals produced by single-copy, targeted GFP in-frame fusions with two different murine Hox genes, Hoxa1 and Hoxc13, are readily detectable by using confocal microscopy. Since Hoxa1 is expressed early and Hoxc13 is expressed late in mouse embryogenesis, this study shows that single-copy GFP gene fusions can be used through most of mouse embryogenesis. Previously, targeted lacZ gene fusions have been very useful for analyzing mouse mutants. Use of GFP gene fusions extends the benefits of targeted lacZ gene fusions by providing the additional utility of a vital marker. Our analysis of the Hoxc13(GFPneo) embryos reveals GFP expression in each of the sites expected from analysis of Hoxc13(lacZneo) embryos. Similarly, Hoxa1(GFPneo) expression was detected in all of the sites predicted from RNA in situ analysis. GFP expression in the foregut pocket of Hoxa1(GFPneo) embryos suggests a role for Hoxa1 in foregut-mediated differentiation of the cardiogenic mesoderm.