Progranulin has multiple functions in several physiological and pathological processes, including embryonic development, wound repair, tumorigenesis, inflammation and neurodegeneration. To investigate the transcriptional regulation of the PGRN gene, a luciferase knock-in reporter system was established in HEK293 cells by integrating luciferase gene in the genome controlled by the endogenous PGRN promoter using CRISPR/Cas9. PCR results demonstrated the site-specific integration of the exogenous luciferase gene into the genome. To validate the novel luciferase knock-in system, a CRISPR/Cas9 transcription activation/repression system for the PGRN gene was constructed and applied to the knock-in system. In addition, phorbol ester (phorbol 12-myristate, 13-acetate), previously reported as activating the expression of PGRN, was applied to the system. The results indicated that luciferase activity was directly correlated with the activity of the PGRN endogenous promoter. This novel system will be a useful tool for investigating the transcriptional regulation of PGRN, and it has great potential in screening the drugs targeting PGRN.

前颗粒蛋白在几种生理和病理过程中具有多种功能，包括胚胎发育，伤口修复，肿瘤发生，炎症和神经变性。为了研究PGRN基因的转录调控，通过使用CRISPR / Cas9将萤光素酶基因整合到由内源PGRN启动子控制的基因组中，在HEK293细胞中建立了萤光素酶敲入报告系统。PCR结果表明外源荧光素酶基因位点特异性整合到基因组中。为了验证新型萤光素酶敲入系统，构建了PGRN基因的CRISPR / Cas9转录激活/抑制系统并将其应用于敲入系统。此外，以前报道为激活PGRN表达的佛波醇酯（佛波醇12-肉豆蔻酸酯，13-乙酸酯）被应用于该系统。结果表明，荧光素酶活性与PGRN内源启动子的活性直接相关。这个新颖的系统将是研究PGRN转录调控的有用工具，并且在筛选靶向PGRN的药物方面具有巨大潜力。