Transcriptomics of Mouse Hepatitis Virus Strain A59 and nsp15 NTD mutants infected C57bl/6 Bone Marrow Derived Macrophages

The study aims to identify non-catalytic residues of Coronavirus nsp15 that regulate EndoU activity during viral replication and to globally evaluate how the loss of EndoU function results in activation of the host innate immune response and subsequent cell death. Total RNA from mock, MHV-A59- and nsp15 mutants-infected C57bl/6 mouse Bone Marrow Derived Macrophages (BMDMs) were extracted in triplicates at 3, 6, 9 and 12 hours post infection (with mock only at 3 hpi). Directional RNA sequencing was performed at the University of Chicago Genomics Facility on a Novaseq X.

本研究旨在鉴定冠状病毒（Coronavirus）非结构蛋白15（nsp15）中在病毒复制过程中调控核糖核酸内切酶U（EndoU）活性的非催化残基，并在全局层面评估EndoU功能缺失如何导致宿主先天免疫应答激活及后续细胞死亡。从空白感染对照组、小鼠肝炎病毒A59株（MHV-A59）感染组及nsp15突变体感染组的C57BL/6小鼠骨髓衍生巨噬细胞（Bone Marrow Derived Macrophages，BMDMs）中提取总RNA，分别于感染后3、6、9、12小时采样（空白对照组仅在感染后3小时采样，hpi即感染后小时（hours post infection）），每份样本设置三次生物学重复。本研究在芝加哥大学基因组学中心（University of Chicago Genomics Facility）使用Novaseq X测序平台完成定向RNA测序。