Phage integrases for the construction and manipulation of transgenic mammals.
Phage integrases for the construction and manipulation of transgenic mammals.
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DOI：
10.1186/1477-7827-1-79
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发表时间：
2003-11-07
期刊：
影响因子：
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通讯作者：
Calos MP
中科院分区：
文献类型：
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作者：
Hollis RP;Stoll SM;Sclimenti CR;Lin J;Chen-Tsai Y;Calos MP
噬菌体整合酶催化两个短att识别位点之间的位点特异性单向重组。当att位点存在于两个不同的DNA分子上时，重组会导致整合，而当att位点位于同一分子上时，重组会导致缺失或倒位。在这里，我们证明了在将供体质粒和整合酶mRNA显微注射到小鼠单细胞胚胎中后，φC31整合酶将DNA整合到小鼠基因组中的内源性序列中的能力。报告了转基因早期胚胎和妊娠中期小鼠。我们还证明了φC31、R4和TP 901 -1噬菌体整合酶重组人类细胞中同一染色体上两个引入的att位点的能力，导致插入物质的缺失。我们比较了这三种整合酶在不同染色体位置催化的哺乳动物染色体缺失的频率。这里回顾的结果介绍这些噬菌体整合酶作为工具的基因组的位点特异性修饰的创建和操作的转基因哺乳动物。
Phage integrases catalyze site-specific, unidirectional recombination between two short att recognition sites. Recombination results in integration when the att sites are present on two different DNA molecules and deletion or inversion when the att sites are on the same molecule. Here we demonstrate the ability of the φC31 integrase to integrate DNA into endogenous sequences in the mouse genome following microinjection of donor plasmid and integrase mRNA into mouse single-cell embryos. Transgenic early embryos and a mid-gestation mouse are reported. We also demonstrate the ability of the φC31, R4, and TP901-1 phage integrases to recombine two introduced att sites on the same chromosome in human cells, resulting in deletion of the intervening material. We compare the frequencies of mammalian chromosomal deletion catalyzed by these three integrases in different chromosomal locations. The results reviewed here introduce these bacteriophage integrases as tools for site-specific modification of the genome for the creation and manipulation of transgenic mammals.