Formation of de novo centromeres and construction of first-generation human artificial microchromosomes
Formation of de novo centromeres and construction of first-generation human artificial microchromosomes
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DOI：
10.1038/ng0497-345
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发表时间：
1997-04-01
期刊：
影响因子：
30.8
通讯作者：
Willard, HF
中科院分区：
文献类型：
--
作者：
Harrington, JJ;VanBokkelen, G;Willard, HF
我们已经结合了长的合成阵列的α卫星DNA与端粒DNA和基因组DNA，以产生人工染色体在人类HT 1080细胞。所得的线性微染色体含有外源性α卫星DNA，在培养中不存在选择的情况下在有丝分裂和细胞遗传学上稳定长达六个月，结合对活性着丝粒特异的着丝粒蛋白，并且估计大小为6-10兆碱基，约为内源性人类染色体大小的五分之一到十分之一。我们的结论是，这种策略的结果在从头着丝粒活性的形成，这样产生的微染色体包含稳定的有丝分裂染色体分离和维护所需的所有序列元素。这种用于构建人类人工染色体的第一代系统应该适合于解剖人类着丝粒的序列要求，以及开发用于治疗应用的构建体。
We have combined long synthetic arrays of alpha satellite DNA with telomeric DNA and genomic DNA to generate artificial chromosomes in human HT1080 cells. The resulting linear microchromosomes contain exogenous alpha satellite DNA, are mitotically and cytogenetically stable in the absence of selection for up to six months in culture, bind centromere proteins specific for active centromeres, and are estimated to be 6-10 megabases in size, approximately one-fifth to one-tenth the size of endogenous human chromosomes. We conclude that this strategy results in the formation of de novo centromere activity and that the microchromosomes so generated contain all of the sequence elements required for stable mitotic chromosome segregation and maintenance. This first-generation system for the construction of human artificial chromosomes should be suitable for dissecting the sequence requirements of human centromeres, as well as developing constructs useful for therapeutic applications.