Photosensitivity and c-GAS-dependent type I IFN activation in lupus patients with TREX1 deficiency

High throughput screening comparing gene expression in human wildtype and patient (TREX1-mutated) fibroblasts, Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to compare NGS-derived retinal transcriptome profiling (RNA-seq) to microarray and quantitative reverse transcription polymerase chain reaction (qRT–PCR) methods and to evaluate protocols for optimal high-throughput data analysis Overall design: RNA sequencing of transcripts isolated out of native and 8h post UV-irradiated cell (fibroblast) samples

本研究为一项高通量筛选实验，对比人类野生型与携带TREX1突变的患者来源成纤维细胞的基因表达差异。下一代测序（Next-generation sequencing, NGS）技术已彻底革新细胞通路的系统性分析范式。本研究的核心目标为：对比基于NGS的视网膜转录组分析（RNA-seq）与基因芯片、定量反转录聚合酶链反应（quantitative reverse transcription polymerase chain reaction, qRT–PCR）技术，并优化适配最优高通量数据分析的实验流程。实验整体设计：对未处理及紫外线照射8小时后的成纤维细胞样本中分离得到的转录本开展RNA测序。