GERM-LINE TRANSMISSION AND EXPRESSION OF A HUMAN-DERIVED YEAST ARTIFICIAL CHROMOSOME
GERM-LINE TRANSMISSION AND EXPRESSION OF A HUMAN-DERIVED YEAST ARTIFICIAL CHROMOSOME
复制标题
DOI：
10.1038/362255a0
复制
发表时间：
1993-03-18
期刊：
影响因子：
64.8
通讯作者：
KLAPHOLZ, S
中科院分区：
文献类型：
--
作者：
JAKOBOVITS, A;MOORE, AL;KLAPHOLZ, S
将数百个千碱基大小的DNA片段引入小鼠胚胎干细胞将极大地提高操纵小鼠基因组的能力。由这种修饰细胞产生的小鼠将可以研究非常大或粗略定位的基因的功能和表达。克隆到酵母人工染色体(YAC)中的大DNA分子是稳定的，并且在Yast1内是可遗传操作的，这表明酵母细胞融合是将大DNA片段转移到哺乳动物细胞的理想方法。通过这种技术将YAC导入不同类型的细胞已有报道2-8；然而，酵母DNA与YAC的结合引起了人们对以这种方式修饰的ES细胞的怀疑。将能够重新克隆小鼠的生殖系5。在这里，我们提供了，据我们所知，第一个种系传递和表达的大的人类DNA片段，通过与酵母球形体融合导入ES细胞。酵母基因组的很大一部分与YAC的共整合不会损害正常的发育。
INTRODUCTION of DNA fragments, hundreds of kilobases in size, into mouse embryonic stem (ES) cells would greatly advance the ability to manipulate the mouse genome. Mice generated from such modified cells would permit investigation of the function and expression of very large or crudely mapped genes. Large DNA molecules cloned into yeast artificial chromosomes (YACs) are stable and genetically manipulable within yeast1, suggesting yeast-cell fusion as an ideal method for transferring large DNA segments into mammalian cells. Introduction of YACs into different cell types by this technique has been reported2-8; however, the incorporation of yeast DNA along with the YAC has raised doubts as to whether ES cells, modified in this way. would be able to recolonize the mouse germ line5. Here we provide, to our knowledge, the first demonstration of germ-line transmission and expression of a large human DNA fragment, introduced into ES cells by fusion with yeast spheroplasts. Proper development was not impaired by the cointegration of a large portion of the yeast genome with the YAC.