The human pre-B cell line Nalm-6 is highly proficient in gene targeting by homologous recombination
The human pre-B cell line Nalm-6 is highly proficient in gene targeting by homologous recombination
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DOI：
10.1089/dna.2006.25.19
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发表时间：
2006-01-01
影响因子：
3.1
通讯作者：
Koyama, H
中科院分区：
文献类型：
--
作者：
Adachi, N;So, S;Koyama, H
基因靶向提供了一个强大的手段来分析基因功能，例如敲除小鼠的研究和最近的工作与高度重组鸡DT 40 B淋巴细胞系。然而，在人类培养的细胞中，基因靶向的低频率是有效产生敲除克隆的严重障碍。此外，常用的人细胞系是核型异常或不稳定的。在这里，我们显示使用启动子无靶向结构，Nalm-6，人前B ALL细胞系，是高度熟练的基因靶向同源重组。事实上，Nalm-6中TP 53基因靶向的效率似乎比HCT 116（一种普遍用于基因靶向的结肠癌细胞系）中的效率高近两个数量级。表达分析显示，在该细胞系中缺乏MSH 2表达。由于Nalm-6具有正常p53状态的稳定的近二倍体核型，我们的结果强调了Nalm-6在人类基因敲除研究中的有用性。
Gene targeting provides a powerful means for analyzing gene function, as exemplified by knockout mouse studies and recent work with the highly recombinogenic chicken DT40 B-lymphocyte line. In human cultured cells, however, the low frequency of gene targeting is a serious barrier to efficiently generate knockout clones. Moreover, commonly used human cell lines are karyotypically abnormal or unstable. Here, we show using promoterless targeting constructs that Nalm-6, a human pre-B ALL cell line, is highly proficient for gene targeting by homologous recombination. Indeed, the efficiency of TP53 gene targeting in Nalm-6 appears nearly two orders of magnitude higher than that in HCT116, a colon cancer cell line popularly used for gene targeting. Expression analysis revealed a lack of MSH2 expression in this cell line. As Nalm-6 has a stable neardiploid karyotype with normal p53 status, our results underscore the usefulness of Nalm-6 for gene knockout studies in humans.