Generation of Stable Drosophila Ovarian Somatic Cell Lines Using the piggyBac System
Generation of Stable Drosophila Ovarian Somatic Cell Lines Using the piggyBac System
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使用piggyBac系统生成稳定的果蝇卵巢体细胞系
DOI：
10.1007/978-1-0716-2380-0_9
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发表时间：
2022
影响因子：
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通讯作者：
Iwasaki Yuka W.
中科院分区：
文献类型：
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作者：
Takeuchi Chikara;Murano Kensaku;Ishikawa Mitsuru;Okano Hideyuki;Iwasaki Yuka W.
转座因子（Transposable elements，TE）在多种生物体的基因组中占有很大比例。因此，抗转座因子机制对于维持基因组的完整性至关重要。PIWI相互作用RNA（piRNA）是抑制果蝇体内TEs的主要力量。卵巢体细胞（OSC），其中核皮尔纳调节是功能性的，已经被用于皮尔纳途径的研究，作为细胞培养系统以阐明皮尔纳途径的分子机制。使用报告系统来监测特定基因的基因调控或过表达的皮尔纳途径的分析将是一种强有力的方法。在这里，我们提出的技术协议，建立稳定的细胞系使用piggyBacsystem，采用的OSCs。这种简单、一致和省时的方案可能会加速对皮尔纳途径的研究。
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 inDrosophilaovaries. 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 thepiggyBacsystem, adopted for OSCs. This easy, consistent, and timesaving protocol may accelerate research on the piRNA pathway.