Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells
Disruption and sequence identification of 2,000 genes in mouse embryonic stem cells
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DOI：
10.1038/33423
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发表时间：
1998-04-09
期刊：
影响因子：
64.8
通讯作者：
Sands, AT
中科院分区：
文献类型：
--
作者：
Zambrowicz, BP;Friedrich, GA;Sands, AT
表达序列标签（EST）（1）和基因组序列形式的序列信息的急剧增加已经产生了“基因功能缺口”，新基因的鉴定速度远远超过了其功能的鉴定速度。通过标记随机诱变在胚胎干细胞（ES）中大规模产生突变的能力为确定哺乳动物系统中的基因功能提供了一种强有力的方法;这种方法在低等生物中得到了很好的建立（2，3）。在这里，我们描述了一种基于基因捕获的高通量诱变方法，该方法允许从突变基因中自动识别序列标签，该方法捕获和突变基因，而不管它们在ES细胞中的表达状态如何。为了便于大规模地研究基因功能，我们使用这些技术来创建称为Omnibank的ES细胞文库，000个基因。
The dramatic increase in sequence information in the form of expressed sequence tags (ESTs)(1) and genomic sequence has created a 'gene function gap', with the identification of new genes far outpacing the rate at which their function can be identified. The ability to create mutations in embryonic stem (ES) cells on a large scale by tagged random mutagenesis provides a powerful approach for determining gene function in a mammalian system; this approach is well established in lower organisms(2,3). Here we describe a high-throughput mutagenesis method based on gene trapping that allows the automated identification of sequence tags from the mutated genes, This method traps and mutates genes regardless of their expression status in ES cells, To facilitate the study of gene function on a large scale, we are using these techniques to create a library of ES cells called Omnibank, from which sequence-tagged mutations in 2,000 genes are described.