Site-specific genomic integration in mammalian cells mediated by phage φC31 integrase
Site-specific genomic integration in mammalian cells mediated by phage φC31 integrase
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DOI：
10.1128/mcb.21.12.3926-3934.2001
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发表时间：
2001-06-01
影响因子：
5.3
通讯作者：
Calos, MP
中科院分区：
文献类型：
--
作者：
Thyagarajan, B;Olivares, EC;Calos, MP
我们之前已经证实噬菌体phi C31整合酶是一种位点特异性重组酶，在人类细胞环境中介导染色体外载体上的attB和attP噬菌体附着位点的有效整合。我们在这里表明，噬菌体attP位点插入到人类和小鼠染色体的不同位置，作为精确的位点特异性整合的有效靶标。此外，我们还描述了人类和小鼠基因组中天然的“伪”attP位点，这些位点也介导了整合酶介导的高效整合。这些位点与attP具有部分序列同一性。这些地点形成了自然发生的整合目标。这种噬菌体整合酶介导的反应代表了一种有效的高级细胞位点特异性整合系统，可能在基因治疗和其他染色体工程策略中具有价值。
We previously established that the phage phi C31 integrase, a site-specific recombinase, mediates efficient integration in the human cell environment at attB and attP phage attachment sites on extrachromosomal vectors. We show here that phage attP sites inserted at various locations in human and mouse chromosomes serve as efficient targets for precise site-specific integration. Moreover, we characterize native "pseudo" attP sites in the human and mouse genomes that also mediate efficient integrase-mediated integration. These sites have partial sequence identity to attP. Such sites form naturally occurring targets for integration. This phage integrase-mediated reaction represents an effective site specific integration system for higher cells and may be of value in gene therapy and other chromosome engineering strategies.