Phage R4 integrase mediates site-specific integration in human cells
Phage R4 integrase mediates site-specific integration in human cells
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DOI：
10.1016/s0378-1119(01)00711-9
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发表时间：
2001-10-31
期刊：
影响因子：
3.5
通讯作者：
Calos, MP
中科院分区：
文献类型：
--
作者：
Olivares, EC;Hollis, RP;Calos, MP
R4整合酶是来源于小链霉菌噬菌体R4基因组的位点特异性单向重组酶。在这里，我们定义紧凑的attB和attP识别位点的R4整合酶和表达的酶在哺乳动物细胞。我们证明，R4整合酶在人类细胞中的功能，进行高效和精确的重组R4 attB和attP位点克隆的染色体外载体。我们还提供的证据表明，酶可以介导整合的传入质粒携带attB或attP网站到内源性序列在人类基因组中。此外，当R4 attB和attP位点通过随机整合或通过使用phi C31整合酶在特定序列处被置于人基因组中时，它们充当携带R4 att位点的引入质粒的整合的靶标。R4整合酶具有作为基因组工程的位点特异性整合工具的直接效用，以及进一步开发的潜力。(C)2001 Elsevier Science B.V.保留所有权利。
The R4 integrase is a site-specific, unidirectional recombinase derived from the genome of phage R4 of Streptomyces parvulus. Here we define compact attB and attP recognition sites for the R4 integrase and express the enzyme in mammalian cells. We demonstrate that R4 integrase functions in human cells, performing efficient and precise recombination between R4 attB and attP sites cloned on an extrachromosomal vector. We also provide evidence that the enzyme can mediate integration of an incoming plasmid bearing an attB or attP site into endogenous sequences in the human genome. Furthermore, when R4 attB and attP sites are placed into the human genome, either by random integration or at a specific sequence by using the phi C31 integrase, they act as targets for integration of incoming plasmids bearing R4 att sites. The R4 integrase has immediate utility as a site-specific integration tool for genome engineering, as well as potential for further development. (C) 2001 Elsevier Science B.V. All rights reserved.