Gene Expression
Nebulas
Gene Expression NebulasSummary: SARS-CoV-2 infection accounts for COVID-19 lung disease and other organ manifestations. Increasing evidence points towards an inflammatory cytokine network as the underlying driver of actual organ damage and severity of the clinical course. Here we show that SARS-CoV-2, like a broad spectrum of other viruses, evokes cellular senescence as a primary stress response in infected cells, among them respiratory epithelial cells, which is – indistinguishably from other forms of cellular senescence – characterized by typical morphological and cell-cycle arrest features, and accompanied by the massive secretion of largely pro-inflammatory cytokines, termed senescence-associated secretory phenotype (SASP).
Overall Design: Examine differentially expressed gene transcripts of senescent cells induced by direct virus infection or conditioned medium from virus-induced senescent cells.
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| Growth Protocol: | A549 and Wi38 cells were cultivated in DMEM supplemented with 10% FBS and Pen-Strep antibiotics; THP-1 cells were cultivated in RPMI-1640 supplemented with 10% FBS and Pen-Strep antibiotics |
| Treatment Protocol: | Cells were infected with virus or cultivated in conditioned medium for 5 days (THP-1: 3 days) and cell pellets were collected for RNA extraction. THP1 VIS samples were split into CD86+ and CD86- fractions using surface labelung with a CD86-biotin antibody and anti-biotin Microbeads from Miltenyi, following the manufacturer´s instructions. |
| Extract Protocol: | Total RNA was extracted using Qiagen RNeasy Plus Mini Kit |
| Library Construction Protocol: | Libraries were prepared by BGI or DKFZ sequencing core facility according to Illumina's TruSeq RNA kit (RS-122-2001) instructions. |
| Molecule Type: | polyA(+) RNA |
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| Library Layout: | SINGLE |
| Library Strand: | -; Forward |
| Platform: | BGISEQ; ILLUMINA |
| Instrument Model: | DNBSEQ-G400; Illumina HiSeq 4000 |
| Strand-Specific: | Unspecific; Specific |
| Data Resource | GEN Sample ID | GEN Dataset ID | Project ID | BioProject ID | Sample ID | Sample Name | BioSample ID | Sample Accession | Experiment Accession | Release Date | Submission Date | Update Date | Species | Race | Ethnicity | Age | Age Unit | Gender | Source Name | Tissue | Cell Type | Cell Subtype | Cell Line | Disease | Disease State | Development Stage | Mutation | Phenotype | Case Detail | Control Detail | Growth Protocol | Treatment Protocol | Extract Protocol | Library Construction Protocol | Molecule Type | Library Layout | Strand-Specific | Library Strand | Spike-In | Strategy | Platform | Instrument Model | Cell Number | Reads Number | Gbases | AvgSpotLen1 | AvgSpotLen2 | Uniq Mapping Rate | Multiple Mapping Rate | Coverage Rate |
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