Use of yeast artificial chromosomes (YACs) in studies of mammalian development: production of beta-globin locus YAC mice carrying human globin developmental mutants.
Use of yeast artificial chromosomes (YACs) in studies of mammalian development: production of beta-globin locus YAC mice carrying human globin developmental mutants.
复制标题
DOI：
10.1073/pnas.92.12.5655
复制
发表时间：
1995-06
影响因子：
11.1
通讯作者：
K. Peterson;Qi Liang Li;C. Clegg;T. Furukawa;P. Navas;E. J. Norton;T. Kimbrough;G. Stamatoyannopoulos
中科院分区：
文献类型：
--
作者：
K. Peterson;Qi Liang Li;C. Clegg;T. Furukawa;P. Navas;E. J. Norton;T. Kimbrough;G. Stamatoyannopoulos
为了测试酵母人工染色体（YAC）是否可用于哺乳动物发育的研究，我们分析了携带两种类型的β-珠蛋白基因座YAC发育突变体的转基因小鼠的表型：（i）在A γ基因的-117位携带G-->A转换的小鼠，这是导致胎儿血红蛋白遗传持续性（HPFH）的希腊A γ形式的原因，和（ii）携带δ-和β-珠蛋白基因缺失的β-珠蛋白基因座YAC转基因系，其5 ′断裂点与缺失型HPFH和δ β-地中海贫血综合征的断裂点相似。携带-117 A γ G->A突变的小鼠显示出延迟的γ-至β-珠蛋白基因转换，并且在发育的成年阶段继续表达A γ-珠蛋白链，如对希腊HPFH携带者所预期的，这表明YAC/转基因小鼠系统允许分析顺式作用基序的发育作用。对携带3'缺失的小鼠的分析首先提供了支持输入增强子负责缺失HPFH表型的假设的证据，其次表明自主沉默是关闭成人中γ-珠蛋白基因的主要机制。总的来说，我们的研究结果表明，携带YAC突变的转基因小鼠提供了一个有用的模型，在发展过程中的基因表达控制的分析。
To test whether yeast artificial chromosomes (YACs) can be used in the investigation of mammalian development, we analyzed the phenotypes of transgenic mice carrying two types of beta-globin locus YAC developmental mutants: (i) mice carrying a G-->A transition at position -117 of the A gamma gene, which is responsible for the Greek A gamma form of hereditary persistence of fetal hemoglobin (HPFH), and (ii) beta-globin locus YAC transgenic lines carrying delta- and beta-globin gene deletions with 5' breakpoints similar to those of deletional HPFH and delta beta-thalassemia syndromes. The mice carrying the -117 A gamma G-->A mutation displayed a delayed gamma- to beta-globin gene switch and continued to express A gamma-globin chains in the adult stage of development as expected for carriers of Greek HPFH, indicating that the YAC/transgenic mouse system allows the analysis of the developmental role of cis-acting motifs. The analysis of mice carrying 3' deletions first provided evidence in support of the hypothesis that imported enhancers are responsible for the phenotypes of deletional HPFH and second indicated that autonomous silencing is the primary mechanism for turning off the gamma-globin genes in the adult. Collectively, our results suggest that transgenic mice carrying YAC mutations provide a useful model for the analysis of the control of gene expression during development.