HSPA6基因对细胞的快速应激和保护具有重要作用。本项目前期，利用CRISPR/Cas9介导的定点整合技术，已成功获得了EGFP与HSPA6同步表达的PK-15细胞系，该细胞系能灵敏的将热、药物等刺激转变为EGFP荧光信号。将该细胞系用于探究猪HSPA6的应激源及其表达调节机制，将会对药/毒物的筛选、病原检测、细胞传感器及畜禽抗逆遗传育种等具有重要意义。接着，本项目首先拟结合荧光显微镜、流式细胞术等方法，将该细胞系用于病原、药/毒物等应激源的系统筛选研究；然后，以特定应激源刺激下细胞荧光强度的变化为依据，通过RNAi、基因定点敲除/突变等探究HSP90、HSP72等因子及启动子调控区对HSPA6表达的影响；最后，再对HSPA6调节因子和序列进行验证和分类。从不同层面对HSPA6基因表达的分子调控机制进行阐明，为全面了解HSPA6基因功能及丰富HSP70家族的生物学功能积淀理论基础。

HSPA6 plays an important role in rapid stress and cytoprotection. In our previous study, CRISPR/Cas9-mediated knock-in strategy was used to successfully obtain a PK-15 cell line that could co-express EGFP and HSPA6. In addition, the cell line could sensitively transfer the stress signal to fluorescent signal from EGFP. Based on the cell line, we speculate that exploring the HSPA6 and its expression regulation mechanism will be of great significance to drug/poison screening, pathogen detection，cell-based sensor and livestock and poultry resistance genetic breeding. First, together with fluorescence microscope and flow cytometry, the cell line will be used for stressor screening such as pathogens and drug/poison. Then, the different fluorescence intensity of the cells that stimulated by specific stressors can be used as a standard for checking HSP90, HSP72 and other factors as well as the regulatory region of the HSPA6 promoter region via site-specific knockout/point mutation strategy. Finally, the relative factors and sequences regulating the gene were confirmed and classified. The molecular regulation mechanism of HSPA6 gene expression can be elucidated from different layers that provide a theoretical basis for totally understanding the function of HSPA6 and enriching biological functions of HSP70 family.