目的：建立由内源性SREBP1启动子控制的荧光素酶敲入报告基因的HEK293细胞系，以研究SREBP1基因的转录调控。
结果：PCR证实了外源荧光素酶基因的单拷贝位点特异性整合到基因组的一个等位基因中，而另一个等位基因中的靶序列缺失了14bp。荧光素酶活性与HEK293-SREBP1-T2A-荧光素酶-KI细胞系中内源性SREBP1基因的启动子活性直接相关。
结论：我们成功地产生了一个新的荧光素酶敲入报告系统，这对于研究SREBP1基因的转录调节和筛选调节SREBP1基因表达的药物或化学分子非常有用。
OBJECTIVE: To establish a HEK293 cell line with a luciferase knock-in reporter controlled by the endogenous
SREBP1 promoter for investigating transcriptional regulation of the
SREBP1 gene.
RESULTS: PCR confirmed the site-specific integration of a single copy of the exogenous luciferase gene into one allele of the genome and a 14 bp deletion of the targeted sequence in the other. Luciferase activity was directly correlated with the promoter activity of the endogenous
SREBP1 gene in the HEK293-
SREBP1-T2A-luciferase-KI cell line cell line.
CONCLUSIONS: We successfully generated a novel luciferase knock-in reporter system, which will be very useful for studying transcriptional regulation of the SREBP1 gene and for screening drugs or chemical molecules that regulate
SREBP1 gene expression.
