Piwi是RNA结合蛋白，在动物的干细胞特性维持、原始生殖细胞（PGC)形成、生殖细胞增殖和分化等方面起重要作用。前期研究表明，青鳉piwi RNA 分布于囊胚期前的每个细胞中，体节期后则定位于PGC、眼和脑；敲降piwi基因影响PGC的增殖和迁移。本研究拟通过RNA荧光原位杂交和免疫组化研究piwi在胚胎和成体的表达模式；通过RNA过表达和敲降研究piwi在胚胎PGC和眼睛发育中的功能；通过 TALENs技术获得piwi敲除的突变体，研究 piwi在成体生殖细胞和眼睛发育中的功能。分析与piwi结合的小RNA（piRNA）；还将通过pull down和免疫共沉淀等技术手段筛选与Piwi蛋白相互作用的基因，揭示Piwi与基因互作的机理。本项目将揭示piwi在生殖细胞和眼睛发育中的功能和作用机理；同时，TALENs技术的建立将为鱼类基因功能研究、遗传育种如抗病品种的培育开拓新的途径。

RNA binding protein Piwi is essential for stem cell maintenance, primordial germ cell (PGC) specification, germ stem cell division and differentiation in diverse organisms. Our previous results showed that medaka piwi RNA was present throughout cleavages stages and widely distributed to every cell until the blastula stage, and concentrated in PGCs, brain and eye during somitogenesis stage; piwi knock down by morpholino (MO) reduced the PGC number and affected PGC migration. This project will continue to study the expression pattern of piwi RNA and protein by fluorescent in situ hybridization and immunohistochemistry. We will further investigate the role of piwi in embryonic PGC and eye development by MO knock down and RNA overexpression. In addition, we will establish piwi null mutant by making use of activator-like effector nucleases (TALENs) mediated genome editing to study the function of piwi in adult germ cells and eye development. Furthermore, we will isolate piwi interacting RNAs (piRNA). Moreover，pull down and immuno-precipitation will be used to screen and identify genes that specifically interact with piwi and the mechanism involved. This study will enhance the understanding of the function and mechanism of piwi in regulating germ cells and eye development, and the establishment of TALENs technique with genome editing will open a new avenue for fish gene function study and the production of food fish with new traits such as virus resistance.