The human beta-globin locus activation region alters the developmental fate of a human fetal globin gene in transgenic mice.
The human beta-globin locus activation region alters the developmental fate of a human fetal globin gene in transgenic mice.
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人β-珠蛋白基因座激活区改变转基因小鼠中人胎儿珠蛋白基因的发育命运。
DOI：
10.1073/pnas.86.18.7033
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发表时间：
1989
影响因子：
11.1
通讯作者：
Stamatoyannopoulos,G
中科院分区：
文献类型：
--
作者：
Enver,T;Ebens,AJ;Forrester,WC;Stamatoyannopoulos,G
我们将含有完整A γ-珠蛋白基因的3.3-内切酶片段连同5'侧翼的1.3个内切酶和3'侧翼DNA的0.37个内切酶连接到含有通常位于人β-珠蛋白基因座5'区的4个发育稳定的超敏位点的2.5-内切酶片段。将该构建体注射到受精小鼠卵中，并在原始和定形红系细胞以及14天胚胎的大脑中分析其表达。所有6个含有完整拷贝的构建体的转基因个体均以红系特异性方式表达转基因。在原始和定形红系细胞中均观察到表达。这与先前使用相同的A γ-珠蛋白基因片段分离的转基因小鼠实验形成鲜明对比，其中表达仅限于原始红系细胞。我们的研究结果表明，在转基因小鼠中，含有发育稳定的珠蛋白基因座超敏位点的区域改变了人胎儿珠蛋白基因的发育阶段特异性。这些观察结果意味着，除了这里使用的序列参与胎儿珠蛋白基因在体内表达的发育控制。在成人红系细胞中表达胎儿珠蛋白的能力允许人们考虑将胎儿珠蛋白基因用于镰状细胞病的基因治疗。
We linked a 3.3-kilobase fragment containing the entire A gamma-globin gene together with 1.3 kilobases of 5' flanking and 0.37 kilobase of 3' flanking DNA to a 2.5-kilobase fragment containing four of the developmentally stable hypersensitive sites normally located in the 5' region of the human beta-globin locus. This construct was injected into fertilized mouse eggs, and its expression was analyzed in the primitive and definitive erythroid cells, as well as the brain of 14-day embryos. All six transgenic individuals that contained intact copies of the construct expressed the transgene in an erythroid-specific fashion. Expression was observed in both primitive and definitive erythroid cells. This is in marked contrast to previous transgenic mice experiments using the same A gamma-globin gene fragment in isolation, where expression was restricted to primitive erythroid cells. Our results show that the region containing the developmentally stable globin locus hypersensitive sites changes the developmental stage specificity of a human fetal globin gene in transgenic mice. These observations imply that sequences additional to those used here are involved in the developmental control of fetal globin gene expression in vivo. The ability to express fetal globin in adult erythroid cells allows one to consider using fetal globin genes for gene therapy of sickle cell disease.