PhiC31 integrase induces efficient site-specific excision in zebrafish.

PhiC31 integrase induces efficient site-specific excision in zebrafish.
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DOI:
10.1007/s11248-010-9394-5
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发表时间:
2011-02
影响因子:
3
通讯作者:
Chen W
Chen W
中科院分区:
生物学4区
文献类型:
--
作者:
Lu J;Maddison LA;Chen W

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定点重组酶催化特定靶点之间的重组,从而高保真地删除、插入、颠倒或交换DNA。除了广泛使用的Cre和FLP重组酶外,来自链霉菌噬菌体的phiC31整合酶系统也可用于真核细胞的这些遗传操作。与Cre和FLP不同,phiC31识别两个异型位点,attB和attP,进行重组。虽然phiC31系统最近已经应用于小鼠和人类细胞系以及果蝇,但它是否也能在斑马鱼中催化有效的DNA重组还没有得到证实。在这里,我们展示了phiC31整合酶有效地诱导斑马鱼胚胎中的外体靶标和染色体靶标的位置特异性缺失。因此,phiC31系统可以用于斑马鱼的遗传操作,扩大了这种脊椎动物模型中可用于遗传操作的工具的库。
Site-specific recombinases catalyze recombination between specific targeting sites to delete, insert, invert, or exchange DNA with high fidelity. In addition to the widely used Cre and Flp recombinases, the phiC31 integrase system from Streptomyces phage may also be used for these genetic manipulations in eukaryotic cells. Unlike Cre and Flp, phiC31 recognizes two heterotypic sites, attB and attP, for recombination. While the phiC31 system has been recently applied in mouse and human cell lines and in Drosophila, it has not been demonstrated whether it can also catalyze efficient DNA recombination in zebrafish. Here we show that phiC31 integrase efficiently induces site-specific deletion of episomal targets as well as chromosomal targets in zebrafish embryos. Thus, the phiC31 system can be used in zebrafish for genetic manipulations, expanding the repertoire of available tools for genetic manipulation in this vertebrate model.
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