受精障碍是导致不孕的重要原因之一，对其深入研究意义重大。前期我们在4名受精障碍患者中鉴定出致病基因WEE2，揭示了其分子机制并通过cRNA靶向治疗成功对一名患者进行分子干预。因此，WEE2 cRNA靶向干预可以作为治疗此类患者的有效方案。但是，目前尚缺乏此类方案的安全性研究。为进行安全性评估，拟在本项目资助下，首次构建WEE2基因敲除受精障碍小鼠模型，运用行为学分析、全基因组测序、质谱分析、免疫荧光、免疫印迹等方法，在体和离体全面评估WEE2 cRNA靶向治疗的安全性。此外，WEE2基因在受精障碍患者中是否存在多种突变类型及相应致病机理尚不清楚。项目将通过靶向测序在受精障碍患者中寻找WEE2基因新的遗传突变，深入和全面地揭示其临床表型和致病机制。该研究一方面将为受精障碍患者的临床诊断提供强有力的分子诊断标记物；另一方面，将为受精障碍患者提供安全有效的临床治疗方案，为未来的精准医疗打下基础。

Fertilization failure is one of the main causes of infertility. It is of great significance to study it. In previous study, we identified the pathogenic gene WEE2 in four patients with fertilization failure, revealed its molecular mechanism and successfully intervened one of the four patients by WEE2 cRNA molecular targeted therapy. Therefore, WEE2 cRNA molecular targeted intervention can be used as an effective treatment for these patients with fertilization failure. However, the safety of WEE2 cRNA targeted therapy remains unknown. In order to evaluate the safety, the project will firstly establish WEE2 knockout transgenic mouse model to access the safety of WEE2 cRNA targeted treatment in vitro and in vivo by the mouse behavioral analysis, whole genome sequencing, mass spectrometry, immunofluorescence, Western Blot et al. In addition, it is not clear whether WEE2 gene has multiple genetic variations in the patients with fertilization failure as well as the related pathogenic mechanism. The project will search for new genetic variations of WEE2 gene by targeted sequencing in the patients with fertilization failure and further reveal the clinical phenotype and pathogenic mechanism. On the one hand, this study will provide a powerful molecular diagnostic marker for the clinical diagnosis of patients with fertilization failure. On the other hand, it will provide a safe and effective solution for the clinical treatment of patients with fertilization failure and lay a foundation for future precision medicine.