Saito Kuniaki
Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.
Methods Mol Biol. 2014;1093:25-33. doi: 10.1007/978-1-62703-694-8_3.
Emerging evidence indicates that PIWI proteins, in collaboration with PIWI-interacting RNAs (piRNAs), play a critical role in retrotransposon silencing in Drosophila gonadal somatic and germ-line cells. The recent establishment of female germ-line stem cells/ovarian somatic sheet and its derivative cell line, ovarian somatic cells (OSCs), allows researchers to study the molecular functions of several protein factors involved in the primary piRNA pathway in Drosophila. Although transgene expression is difficult to achieve in gonad-derived cell lines, transfection of both expression vectors and knockdown reagents is highly effective in OSCs. Here, I focus on techniques that knockdown or overexpress genes of interest in OSCs.
新出现的证据表明,PIWI蛋白与PIWI相互作用RNA(piRNA)协同作用,在果蝇性腺体细胞和生殖细胞的逆转座子沉默中起关键作用。最近建立的雌性生殖系干细胞/卵巢体细胞片及其衍生细胞系——卵巢体细胞(OSC),使研究人员能够研究果蝇初级piRNA途径中几种蛋白质因子的分子功能。尽管在性腺来源的细胞系中难以实现转基因表达,但在OSC中,表达载体和敲低试剂的转染都非常有效。在这里,我重点介绍在OSC中敲低或过表达感兴趣基因的技术。