Illuminating DNA replication during Drosophila development using TALE-lights.
Illuminating DNA replication during Drosophila development using TALE-lights.
复制标题
使用 TALE 灯照亮果蝇发育过程中的 DNA 复制。
DOI：
10.1016/j.cub.2014.01.023
复制
发表时间：
2014
期刊：
影响因子：
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通讯作者：
O'Farrell,PatrickH
中科院分区：
文献类型：
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作者：
Yuan,Kai;Shermoen,AntonyW;O'Farrell,PatrickH
新发现允许使用类转录激活因子效应器 (TALE) 的模块化识别单元或 CRISPR 的指导 RNA 进行 DNA 序列识别的系统工程。工程化的特异性提供了引导特定序列的广泛活性的潜力[1-3]，并且靶向核酸酶引起定向诱变[4-6]。在这里，我们使用称为 TALE 光的荧光融合来标记序列，并使用它们来跟踪果蝇主要胚胎转变期间特定卫星序列的复制。我们表明可以测量单个序列的复制时间。随着胚胎的发育，细胞周期延长，卫星序列的复制时间在更长的 S 期内转移到较晚的时间。在这里，我们表明卫星序列的紧凑焦点随着它们的复制而扩展，并且卫星序列（359-bp）在中囊胚转变（MBT）后显示出特别明显的细胞周期后期复制的转变。这种复制行为表明发育信号可以分别影响不同卫星序列的时间。
New discoveries allow systematic engineering of DNA sequence recognition using the modular recognition units of the transcription activator-like effectors (TALEs) or the guide RNA of the CRISPRs. The engineered specificity offers the potential to guide a wide range of activities to particular sequences [1–3], and targeted nucleases cause directed mutagenesis [4–6]. Here we have tagged sequences using fluorescent fusions that we call TALE-lights, and have used these to follow replication of particular satellite sequences during a major embryonic transition inDrosophila. We show that replication-timing of individual sequences can be measured. As embryos develop, the cell cycle extends and the timing of replication of satellite sequences shifts to later times within the more prolonged S phase. Here we show that the compact foci of satellite sequences expand in conjunction with their replication and that a satellite sequence, 359-bp, shows a particularly marked shift to later replication in the cell cycle after the midblastula transition (MBT). This replication behavior suggests that developmental signals can separately influence the timing of different satellite sequences.