GENE TARGETING AT THE HUMAN CD4 LOCUS BY EPITOPE ADDITION
GENE TARGETING AT THE HUMAN CD4 LOCUS BY EPITOPE ADDITION
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DOI：
10.1101/gad.4.2.157
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发表时间：
1990-02-01
影响因子：
10.5
通讯作者：
BERG, P
中科院分区：
文献类型：
--
作者：
JASIN, M;ELLEDGE, SJ;BERG, P
在人类T细胞系中，通过一种被称为表位加成的方法，已经实现了在CD4基因座上的同源重组。内源性的CD4基因为残缺的Thy-1基因提供转录、翻译和前导序列，导致小鼠Thy-1表位在人类细胞表面的表达。用荧光激活细胞分选仪(FACS)筛选THY-1+细胞。据估计，同源与非同源整合事件的浓缩倍数为700倍，因此70%的Thy-1阳性细胞来自于基因靶向。Thy-1+细胞系中有三个只表达目标等位基因的蛋白质；因此，这些细胞在功能上是CD4-。
Homologous recombination at the CD4 locus in a human T-cell line has been achieved by an approach called epitope addition. The endogenous CD4 gene provided transcription, translation, and leader sequences to a crippled introduced Thy-1 gene, resulting in the expression of murine Thy-1 epitopes on the surface of the human cells. Thy-1+ cells were selected using the Fluorescence Activated Cell Sorter (FACS). An estimated 700-fold enrichment for homologous versus nonhomologous integration events was obtained, such that 70% of cells scoring positive for Thy-1 were derived from gene targeting. Three of the Thy-1+ cell lines expressed protein only from the targeted allele; thus, these cells were functionally CD4-.