Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.
Department of Physiology, Keio University School of Medicine, Tokyo, Japan.
Methods Mol Biol. 2022;2509:143-153. doi: 10.1007/978-1-0716-2380-0_9.
Transposable elements (TEs) constitute a large proportion of the genome in multiple organisms. Therefore, anti-transposable element machineries are essential to maintain genomic integrity. PIWI-interacting RNAs (piRNAs) are a major force to repress TEs in Drosophila ovaries. Ovarian somatic cells (OSC), in which nuclear piRNA regulation is functional, have been used for research on piRNA pathway as a cell culture system to elucidate the molecular mechanisms underlying the piRNA pathway. Analysis of piRNA pathway using a reporter system to monitor the gene regulation or overexpression of specific genes would be a powerful approach. Here, we present the technical protocol to establish stable cell lines using the piggyBac system, adopted for OSCs. This easy, consistent, and timesaving protocol may accelerate research on the piRNA pathway.
转座元件 (TEs) 在多种生物体的基因组中构成了很大的比例。因此,抗转座元件机制对于维持基因组完整性是必不可少的。PIWI 相互作用 RNA (piRNAs) 是抑制果蝇卵巢中转座元件的主要力量。卵巢体细胞 (OSC) 中核 piRNA 调节功能正常,已被用作研究 piRNA 途径的细胞培养系统,以阐明 piRNA 途径的分子机制。使用报告系统分析 piRNA 途径以监测特定基因的基因调控或过表达将是一种强有力的方法。在这里,我们介绍了使用 piggyBac 系统建立稳定细胞系的技术方案,该系统适用于 OSCs。这种简单、一致且节省时间的方案可能会加速 piRNA 途径的研究。