RNA干扰是研究未知蛋白在细胞分裂过程中功能的最重要手段之一。在体细胞有丝分裂的研究中，研究者可用脂质体介导的siRNA、慢病毒介导的shRNA等基因敲降方法进行 "全蛋白组"或"局域蛋白组"式的大规模表型筛选。而在卵母细胞减数分裂的研究中，由于卵母细胞透明带的阻隔，这些手段都不能产生有效的基因敲降，因此只能用效率低、易损伤卵的显微注射法进行。本申请在成功的前期试验（12种蛋白）的基础上，提出用非脂质体的新型纳米颗粒介导的siRNA基因敲降来进行扩大规模（80种蛋白）的减数分裂相关蛋白的筛选和功能鉴定，一方面旨在检验这种高效、简便、无损伤的技术在更大规模的蛋白功能筛选中是否依然稳定可靠，为将来可能的多单位协作进行"全蛋白组"规模研究卵母细胞减数分裂中特定事件相关的所有未知蛋白奠定基础；另一方面随着研究的进展将该技术介绍给国内同行，实现协作共赢，从而为我国生殖医学研究的进一步提升贡献力量。

RNA interference is one of the most important tools to study the function of unknown proteins in cell division. In mitosis of somatic cells, researchers can use liposome-mediated siRNA transfection or lentivirus-mediated shRNA gene knockdown to achieve phenotypic screen at "whole proteome" or "partial proteome" scale. While in meiosis of oocytes, by these means researchers couldn't generate efficient gene knockdown due to the blockage of pellucid zone, so they have to use microinjection, which is inefficient and harmful. In our proposal, based on previous successful trial at small scale (12 proteins), we plan to use novel non-liposomal nanoparticle-mediated siRNA gene knockdown to screen and functionally identify meiosis-related unknown proteins at larger scale (100 proteins). Through this, on the one hand we can testify whether this efficient, simple, harmless technique are still stable and reliable at a larger scale, thereby lay the foundation for the possible multi-institute collaborative study on the function of all the proteins in a certain meiotic process at "whole proteome" scale; on the other hand introduce it to other researchers so that we can go forward together and contribute to the progress of reproductive medicine in China.