Transcriptional profiling of A549 cells infected with high capacity MCMV vector.

To gain a detailed knowledge about the virus cycle of murine cytomegalovirus (MCMV) and its high capacity vector in cross-species settings, we analyzed the early and late viral and host transcriptome upon infection of human cells. ARPE-19, A549, and 911 cells (5.0×105 cells/well) were infected in a 24-well format 4 h post-seeding with MCMV-wt or Q4-LRBAs-GLuc, a high capacity replication competent vector based on MCMV, at an MOI of 3. As control, we used infection of mouse embryonic fibroblast, which is the natural host of the wild type MCMV, treated similarly. Harvesting occurred at 8 hpi and 31 hpi through centrifugation at 1.000 g for 5 min. Cell pellets were washed with PBS, re-suspended in 350 µL of RLT buffer, and processed using the RNeasy Mini kit as per the manufacturer’s instructions in independent triplicates. At least 1.500 ng of RNA in a 30 µL volume was isolated and sent for Illumina next-generation sequencing, resulting in paired-end sequencing with a read length of 2×100 bp and a depth of 20 million reads. This submission contains the reads, we obtained analyzing the infections of A549 cells. The data sheet for the samples and the reference genomes (.gb), which we used in the analysis published in Riedl et al. Vaccines 2024 can be found in the REFERENCES_A549.zip folder. Please, find the control reads for uninfected cells in a separate upload entitled: Mock infected controls for transcriptional profiling of infections with high capacity MCMV vector (DOI 10.5281/zenodo.1250410).为深入解析鼠巨细胞病毒（murine cytomegalovirus, MCMV）及其基于该病毒构建的高容量复制型载体在跨物种环境中的病毒增殖周期，我们对感染后人源细胞的病毒与宿主转录组早晚期表达谱进行了系统性分析。实验选用ARPE-19、A549及911细胞，以每孔5.0×10^5个细胞的密度接种于24孔板，接种4小时后以感染复数（multiplicity of infection, MOI）为3的剂量，分别用野生型MCMV（MCMV-wt）或基于MCMV构建的高容量复制型载体Q4-LRBAs-GLuc进行感染。作为对照，我们采用野生型MCMV的天然宿主——小鼠胚胎成纤维细胞开展同等条件的感染处理。分别于感染后8小时（8 hpi）与31小时（31 hpi）收集细胞，通过1000 g离心5分钟富集细胞沉淀。用磷酸盐缓冲液（phosphate-buffered saline, PBS）洗涤细胞沉淀后，将其重悬于350 μL RLT缓冲液中，严格按照试剂盒说明书使用RNeasy Mini试剂盒完成独立三次重复的RNA提取。最终从每份30 μL的体系中分离得到至少1500 ng总RNA，送至Illumina平台开展下一代测序，采用双端测序策略，测序读长为2×100 bp，测序深度达2000万条reads。本提交内容包含我们对A549细胞感染实验的测序reads数据。用于本分析的样本信息表、参考基因组文件（.gb格式）已收录于Riedl等人2024年发表于《Vaccines》的研究中，相关文件可在REFERENCES_A549.zip压缩包中获取。未感染细胞的对照reads数据已单独上传，标题为：《高容量MCMV载体感染转录组分析的Mock感染对照》（DOI 10.5281/zenodo.1250410）。