Group Fungal Epigenomics, Department of Mycology, Institut Pasteur, Paris 75015, France.
Group Fungal Epigenomics, Department of Mycology, Institut Pasteur, Paris 75015, France.
Fungal Genet Biol. 2020 Mar;136:103316. doi: 10.1016/j.fgb.2019.103316. Epub 2019 Dec 9.
The development of a tetO/TetR system in the fungus Neurospora crassa is described. The system includes (i) a synthetic gene encoding a TetR variant fused to GFP, and (ii) a standard tetO array integrated homologously, as a proof of principle, near the his-3 gene. The localization of TetR-GFP at the tetO array (observed by fluorescence microscopy) can be disrupted by the application of tetracycline. The full-length array is stable during vegetative growth, but it triggers strong repeat-induced point mutation (RIP) by the RID-dependent as well as the DIM-2-dependent pathways during the sexual phase. Thus, both RIP pathways must be inactivated to allow the faithful inheritance of the unmodified construct. In summary, this study introduces a new molecular tool into Neurospora research, and suggests that the standard tetO array can self-engage in recombination-independent homologous pairing.
文中描述了在真菌粗糙脉孢菌中 tetO/TetR 系统的发展。该系统包括 (i) 一个编码融合 GFP 的 TetR 变体的合成基因，以及 (ii) 一个标准的 tetO 阵列，作为同源整合的原理证明，靠近 his-3 基因。tetracycline 的应用可破坏 TetR-GFP 在 tetO 阵列上的定位（通过荧光显微镜观察）。在营养生长期间，完整的阵列是稳定的，但它在有性阶段通过 RID 依赖和 DIM-2 依赖途径触发强烈的重复诱导点突变 (RIP)。因此，必须使两种 RIP 途径失活，以允许未修饰的构建体的忠实遗传。总之，这项研究将一种新的分子工具引入了 Neurospora 研究，并表明标准的 tetO 阵列可以自行参与重组非依赖性同源配对。
