RECOMBINASE-MEDIATED GENE ACTIVATION AND SITE-SPECIFIC INTEGRATION IN MAMMALIAN-CELLS
RECOMBINASE-MEDIATED GENE ACTIVATION AND SITE-SPECIFIC INTEGRATION IN MAMMALIAN-CELLS
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DOI：
10.1126/science.1900642
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发表时间：
1991-03-15
期刊：
影响因子：
56.9
通讯作者：
WAHL, GM
中科院分区：
文献类型：
--
作者：
OGORMAN, S;FOX, DT;WAHL, GM
在哺乳动物细胞中实施了基于由来自酵母的FLP重组酶介导的转染序列的条件重组的基因激活和位点特异性整合的二元系统。 在几个细胞系中，FLP快速而精确地重组其特定靶序列的拷贝，以激活一个原本沉默的β-半乳糖苷酶报告基因。 标记细胞的克隆通过在沉默报告基因的染色体整合拷贝内的切除重组产生。 通过逆反应，转染的DNA的整合被靶向到特定的染色体位点。 结果表明，FLP可用于嵌合激活或标记转基因用于脊椎动物发育分析，并有效地整合转染的DNA在预定的染色体位置。
A binary system for gene activation and site-specific integration, based on the conditional recombination of transfected sequences mediated by the FLP recombinase from yeast, was implemented in mammalian cells. In several cell lines, FLP rapidly and precisely recombined copies of its specific target sequence to activate an otherwise silent beta-galactosidase reporter gene. Clones of marked cells were generated by excisional recombination within a chromosomally integrated copy of the silent reporter. By the reverse reaction, integration of transfected DNA was targeted to a specific chromosomal site. The results suggest that FLP could be used to mosaically activate or inactivate transgenes for analysis of vertebrate development, and to efficiently integrate transfected DNA at predetermined chromosomal locations.