Low level of Hox1.3 gene expression does not preclude the use of promoterless vectors to generate a targeted gene disruption. off.
Low level of Hox1.3 gene expression does not preclude the use of promoterless vectors to generate a targeted gene disruption. off.
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Hox1.3 基因表达水平低并不排除使用无启动子载体来产生靶向基因破坏。
DOI：
10.1128/mcb.11.11.5578-5585.1991
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发表时间：
1991
影响因子：
5.3
通讯作者：
Robertson,EJ
中科院分区：
文献类型：
--
作者：
Jeannotte,L;Ruiz,JC;Robertson,EJ
最近设计了多种实验方法通过同源重组来突变哺乳动物基因。在本报告中，我们描述了使用其中两种方法对 Hox1.3 基因座的破坏，即正负选择和无启动子基因的激活。有趣的是，我们观察到这两种程序的针对性破坏频率相似。鉴于用于这些研究的胚胎干细胞系中 Hox1.3 表达水平极低，无启动子载体的高频率靶向破坏是出乎意料的。这些数据表明，需要靶基因的最小表达来富集无启动子载体的同源重组事件，从而增强无启动子基因方法作为通过同源重组突变哺乳动物基因的一般策略。* †
A variety of experimental approaches have been devised recently to mutate mammalian genes by homologous recombination. In this report, we describe the disruption of the Hox1.3 locus by using two of these approaches, namely, positive-negative selection and activation of a promoterless gene. Interestingly, we observe similarly high frequencies of targeted disruption with both procedures. The high frequency of targeted disruption with a promoterless vector was unexpected given the extremely low level of Hox1.3 expression in the embryonic stem cell line used for these studies. These data indicate that minimal expression of the target gene is required to enrich for homologous recombination events with promoterless vectors and thus enhance the promoterless gene approach as a general strategy to mutate mammalian genes by homologous recombination.* †