An induction gene trap screen in embryonic stem cells: Identification of genes that respond to retinoic acid in vitro
An induction gene trap screen in embryonic stem cells: Identification of genes that respond to retinoic acid in vitro
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DOI：
10.1073/pnas.93.4.1677
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发表时间：
1996-02-20
影响因子：
11.1
通讯作者：
Wurst, W
中科院分区：
文献类型：
--
作者：
Forrester, LM;Nagy, A;Wurst, W
我们开发了一种新的诱导基因陷阱方法，该方法在体外预先选择整合到受体/配体介导的信号传导通路下游的基因中。使用这种方法，我们在胚胎干细胞中鉴定了20个基因陷阱整合，其中9个被诱导，11个在暴露于外源性视黄酸（RA）后被抑制。所有这些整合，但一个显示出独特的空间限制或组织特异性的表达模式之间的8.5和11.5天的胚胎发生。有趣的是，在胚胎发生期间受RA水平改变影响的组织中观察到表达。从六个整合的融合转录本的序列分析发现五个新的基因序列和先前确定的原癌基因c-fyn。迄今为止，生殖系传播和育种已经发现了一个纯合胚胎致死和三个纯合可行的插入，这些研究表明，这种诱导基因陷阱方法用于识别和突变信号转导通路下游基因的潜力。
We have developed a novel induction gene trap approach that preselects in vitro for integrations into genes that lie downstream of receptor/ligand-mediated signaling pathways, Using this approach, we have identified 20 gene trap integrations in embryonic stem cells, 9 of which were induced and 11 of which were repressed after exposure to exogenous retinoic acid (RA). All but one of these integrations showed unique spatially restricted or tissue-specific patterns of expression between 8.5 and 11.5 days of embryogenesis. Interestingly, expression was observed in tissues that are affected by alterations in RA levels during embryogenesis. Sequence analysis of fusion transcripts from six integrations revealed five novel gene sequences and the previously identified protooncogene c-fyn. To date, germ-line transmission and breeding has uncovered one homozygous embryonic lethal and three homozygous viable insertions, These studies demonstrate the potential of this induction gene trap approach for identifying and mutating genes downstream of signal transduction pathways.