Hippo通路在调控器官生长、细胞增殖和凋亡中发挥重要作用，我们发现Brahma与Yki-Sd形成复合物，调节Hippo信号通路（Cell.Signal，2015）。miRNA作为重要调节因子可以与Hippo通路交互作用，参与细胞增殖与凋亡的调节，相关作用机制仍需深入研究。我们前期在果蝇中筛选到一个新的Hippo通路调节miRNA：miR-285。发现miR-285下调Hippo通路多个下游基因表达，促进细胞凋亡，导致发育异常。本项目旨在此基础上深入研究miR-285通过Hippo通路调节细胞增殖与凋亡的作用机制及其对发育的生理意义。拟通过鉴定miR-285靶基因，明确miR-285调节Hippo通路的分子机制；利用CRISPR/Cas9等技术构建miR-285突变果蝇，检测其细胞增殖、凋亡及发育的异常，探讨发育中miR-285的生理功能，为深入理解Hippo通路的调控网络提供理论依据。

Hippo pathway is a highly conserved signaling network that controls organ size by restricting cell proliferation and promoting apoptosis. Our previous research demonstrated that Brahma regulated the Hippo pathway activity through forming complex with Yki-Sd (Cell.Signal, 2015). microRNAs have emerged as key regulators of cellular proliferation and apoptosis by cross-talking with Hippo pathway. However, many crucial questions await answers before the regulatory mechanism can be fully elucidated. Recently we identified miR-285 as a novel microRNA gene in Drosophila that can regulate Hippo signaling activity. Our preliminary data showed that overexpression of miR-285 in Drosophila resulted in a strong undergrowth phenotype. Furthermore, we found that overexpression of miR-285 significantly downregulate multiple downstream target genes of Hippo signaling pathway and promote apoptosis. In this project we will further study the miR-285 mediated regulatory mechanism of cellular proliferation and apoptosis through Hippo pathway. We will identify the direct target genes of miR-285 by utilizing Drosophila RNAi libraries and reporter gene system based on bioinformatic analysis. Moreover we will explore the biological role of miR-285 during Drosophila development by generating miR-285 mutant fly using CRISPR/Cas9 and imprecise transposon excision technique. This study will enrich our understanding of the regulatory network of Hippo pathway during development.