Effective gene silencing in Drosophila ovarian germline by artificial microRNAs
Effective gene silencing in Drosophila ovarian germline by artificial microRNAs
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DOI：
10.1038/cr.2011.44
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发表时间：
2011-04
期刊：
影响因子：
44.1
通讯作者：
Hailong Wang;Yanjun Mu;Dahua Chen
中科院分区：
文献类型：
--
作者：
Hailong Wang;Yanjun Mu;Dahua Chen
Drosophila oogenesis is of great interest because it represents an excellent model system to study a number of fascinating biological processes, such as stem cell regulation, germ cell meiosis and oocyte determination, as well as signal interactions between germline and soma. A typical Drosophila ovary is composed of 16-20 ovarioles, each consisting of an anterior functional unit called a germarium and a linear string of differentiated egg chambers posterior to the germarium [1](Figure 1A and 1B). Drosophila oogenesis initiates at the tip of the germarium, when a germline stem cell (GSC) divides asymmetrically to generate a daughter GSC and a cystoblast that eventually develops into a mature egg [2](Figure 1C and 1E). During the last decades, much progress has been made on identifying the key regulators controlling oogenesis, using traditional forward genetic screening via ethyl methanesulfonate and P-element mutagenesis. Such approaches are undoubtedly fruitful. To date, many sterile mutations that disrupt genes necessary for oogenesis have been generated. However, systematic genetic screening and subsequent identification of the role of genes in oogenesis largely depend on phenotypic analysis; therefore, the effects of many unidentified genes on oogenesis are often masked by lethality. Thus, given the limitation of inherent bias in traditional forward genetics, it would be advantageous to develop alternative methods to remove a gene’s function in a tissue-specific manner for screening and characterization of novel gene functions in oogenesis.Small RNA-mediated gene silencing controls a wide range of processes during development. So far, three classes of small RNAs have been characterized in Drosophila, including small interfering RNAs (siRNAs), microRNAs (miRNAs) and Piwi-interacting RNAs [3]. Double-stranded siRNA-mediated gene silencing, as a potent tool, has been widely used for knocking down gene expression in S2 cell cultures and in multiple tissues in flies. In Drosophila oogenesis, the siRNA pathway is apparently activated during egg activation,
