Gene expression profiling of dsDNA-transfected reconstituted mouse embryonic fibroblasts

Analysis of dsDNA-induced innate immune response at gene expression level. The hypothesis tested in this study was that K224R mutation significantly inhibits hSTING activity. Results provide important information of the response of hSTING and its variants to cytosolic dsDNA, such as induction of type I IFN genes and proinflammatory cytokine genes. Sting-/- MEFs were reconstituted with wild type hSTING, hSTING-K224R or hSTING-K289R. Total RNA obtained from dsDNA-transfected MEFs was compared to mock-transfected MEFs.

本研究开展了基于基因表达水平的双链DNA（double-stranded DNA，dsDNA）诱导先天免疫应答分析。本研究所验证的科学假说为：K224R突变可显著抑制人干扰素基因刺激因子（human STING，hSTING）的活性。研究结果为阐明hSTING及其变体对胞质dsDNA的应答机制提供了重要依据，具体涵盖I型干扰素（type I IFN）基因与促炎细胞因子基因的诱导表达情况。本研究将野生型hSTING、hSTING-K224R或hSTING-K289R重组导入STING基因敲除的小鼠胚胎成纤维细胞（Mouse Embryonic Fibroblasts，MEFs）中；将经dsDNA转染的MEFs提取的总RNA，与空白转染的MEFs提取的总RNA进行对照比较。