Expression data from dsDNA-stimulated mouse embryonic fibroblasts. Mus musculus

Transfection of dsDNA into many mammalian cell types indues the production of type I interferons and interferon-stimulated genes. We performed an siRNA screen to identify genes involved in this innate immune response, and identified Abcf1. We used microarrays to determine which genes are regulated by ABCF1 following dsDNA stimulation. Overall design: We treated p53-/- mouse embryonic fibroblasts (MEFs) with siRNAs targeting Abcf1 or Irf3 (positive control) or a negative control siRNA (siCtrl), and then stimulated the cells with 45-base pair dsDNA for 6 hrs or left the cells unstimulated. We then lysed the cells and hybridized the RNA to Affymetrix microarrays.

双链DNA（dsDNA）转染至多种哺乳动物细胞类型，可诱导I型干扰素（type I interferons）及干扰素刺激基因（interferon-stimulated genes）的产生。本研究开展小干扰RNA（siRNA）筛选以鉴定参与该天然免疫应答的基因，并成功鉴定出ATP结合盒式蛋白F1（Abcf1）。为探究双链DNA（dsDNA）刺激后受ABCF1调控的基因表达谱，我们采用基因芯片（microarrays）技术进行检测。

实验设计总览：我们将p53基因敲除的小鼠胚胎成纤维细胞（p53-/- mouse embryonic fibroblasts, MEFs）分别用靶向Abcf1的siRNA、靶向干扰素调节因子3（Irf3，阳性对照）的siRNA，以及阴性对照siRNA（siCtrl）进行处理；随后用45碱基对的双链DNA刺激细胞6小时，同时设置未刺激对照组。收集细胞并裂解后，提取RNA与Affymetrix基因芯片进行杂交实验。