Vollum Institute, OHSU, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA.
Transgenic Res. 2011 Feb;20(1):183-9. doi: 10.1007/s11248-010-9394-5. Epub 2010 Jun 17.
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.
位点特异性重组酶能在特定的靶向位点之间催化高保真的 DNA 切除、插入、反转或交换。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中的这些遗传操作。与 Cre 和 Flp 不同,phiC31 识别两个异型位点 attB 和 attP 以进行重组。虽然 phiC31 系统最近已应用于小鼠和人类细胞系以及果蝇中,但尚未证明它是否也能在斑马鱼中催化有效的 DNA 重组。本文中我们展示了 phiC31 整合酶能有效地诱导斑马鱼胚胎中外源和染色体靶标上的特异性缺失。因此,phiC31 系统可用于斑马鱼中的遗传操作,扩展了该脊椎动物模型中遗传操作可用工具的范围。