HPRT minigene generates chimeric transcripts as a by-product of gene targeting
HPRT minigene generates chimeric transcripts as a by-product of gene targeting
复制标题
DOI：
10.1002/dvg.20300
复制
发表时间：
2007-05-01
期刊：
影响因子：
1.5
通讯作者：
Hasty, Paul
中科院分区：
文献类型：
--
作者：
Holcomb, Valerie B.;Kim, Tae Moon;Hasty, Paul
HPRT小基因是用于在小鼠胚胎干（ES）细胞中进行基因靶向的选择盒，并且它是独特的，因为可以针对其存在和不存在而应用选择。这个小基因有两个外显子簇，由一个小的内含子和剪接序列分开。我们发现这些外显子簇剪接到靶基因的外显子中，形成两种不同类型的嵌合转录本。第一类由内源启动子表达，包括剪接成小基因外显子3-8的上游靶基因外显子。第二类由小基因的PGK启动子表达，包括剪接到下游靶基因外显子中的小基因外显子1-2。这些嵌合转录物可以产生影响表型的嵌合蛋白。因此，我们设计了两个floxed HPRT小基因，其允许通过Cre介导的重组去除小基因的5'半或整个小基因。
The HPRT minigene is a selection cassette used for gene targeting in mouse embryonic stem (ES) cells and, it is unique since selection may be applied for its presence and absence. This minigene has two exon clusters separated by a small intron and splicing sequences. We find these exon clusters splice into exons from the target gene forming two different classes of chimeric transcripts. The first class is expressed by the endogenous promoter and includes upstream target gene exons spliced into minigene exons 3-8. The second class is expressed by the mini-gene's PGK promoter and includes minigene exons 1-2 spliced into downstream target gene exons. These chimeric transcripts may produce chimeric proteins that could influence phenotype. Therefore, we have designed two floxed HPRT minigenes that permit removal of either the 5' half of the minigene or the entire minigene via Cre-mediated recombination.