Identification of immunogenic double-stranded RNAs in human cell lines

We expressed different isoforms of ADAR proteins in human cells without A-to-I RNA editing activity to test which isoform(s) was able to rescue the interferon induction phenotypes caused by activation of MDA5 in the absense of double-stranded RNA editing. Using the rescue cells, we sequenced the RNA and profiled RNA editing in order to identify the key RNA editing substrates for MDA5 sensing. Overall design: Total RNA profiles of wild type (WT) and mutant HEK293 cells expressing different ADAR isoforms/proteins were generated by deep sequencing

我们在丧失A-to-I RNA编辑（A-to-I RNA editing）活性的人类细胞中表达不同亚型的ADAR（腺苷脱氨酶作用于RNA，Adenosine Deaminase Acting on RNA）蛋白，以探究何种亚型能够挽救因双链RNA（double-stranded RNA）编辑缺失导致MDA5（黑素瘤分化相关蛋白5，Melanoma Differentiation-Associated protein 5）激活所引发的干扰素诱导表型。我们利用该挽救细胞系对RNA进行测序，并开展RNA编辑谱分析，以鉴定MDA5感知过程中的关键RNA编辑底物。实验整体设计：通过深度测序（deep sequencing）构建表达不同ADAR亚型/蛋白的野生型（wild type, WT）与突变型HEK293细胞的总RNA表达谱。