Gene Expression
Nebulas
Gene Expression NebulasSummary: The development of tractable animal models that faithfully reproduce COVID-19 pathogenesis would arguably be a major benefit. Infection of transgenic mice expressing the human version of the SARS-CoV-2 receptor (hACE2) by intranasal instillation of a liquid SARS-CoV-2 suspension under deep anesthesia results in disproportionate high CNS infection leading to fatal encephalitis, which is rarely observed in humans and severely limits this model’s usefulness. Here, we describe the characterization of a novel COVID-19 mouse model based on an inhalation tower system that allows to expose unanesthetized mice to aerosolized virus under controlled conditions. Aerosol exposure of K18-hACE2 transgenic mice to SARS-CoV-2 resulted in robust viral replication in the respiratory tract, anosmia, and airway obstruction, but did not lead to fatal viral neuroinvasion. When compared to intranasal inoculation, aerosol infection resulted in a more pronounced lung pathology including increased immune infiltration, fibrin deposition and a transcriptional signature comparable to that observed in humans. This model may prove useful for studies of viral transmission, disease pathogenesis (including long-term consequences of SARS-CoV-2 infection) and therapeutic interventions.
Overall Design: Bulk RNA-seq of lung homogenates 6 days after SARS-CoV-2 infection. Comparison between intranasal- (n=3) and aerosol- (n=4) infected mice, 2 PBS-treated mice are added as control
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| Extract Protocol: | 100 ml of Chloroform were added in 500 ml of homogenized sample and total RNA was extracted using ReliaPrep™ RNA Tissue Miniprep column (Promega, Cat #Z6111). Total RNA was isolated according to the manufacturer’s instructions. qPCR was performed using TaqMan Fast virus 1 Step PCR Master Mix (Lifetechnologies #4444434), standard curve was drawn with 2019_nCOV_N Positive control (IDT#10006625), primer used are: 2019-nCoV_N1- Forward Primer (5’-GAC CCC AAA ATC AGC GAA AT-3’), 2019-nCoV_N1- Reverse Primer (5’-TCT GGT TAC TGC CAG TTG AAT CTG-3’) 2019-nCoV_N1-Probe (5’-FAM-ACC CCG CAT TAC GTT TGG TGG ACC-BHQ1-3’) (Centers for Disease Control and Prevention (CDC) Atlanta, GA 30333) |
| Library Construction Protocol: | Libraries were purified with AMPure XP beads, quantified using Qubit 3.0 and single-end sequenced (75 bp) on an Illumina NextSeq 500. |
| Molecule Type: | rRNA- RNA |
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| Library Layout: | SINGLE |
| Library Strand: | - |
| Platform: | ILLUMINA |
| Instrument Model: | Illumina NextSeq 500 |
| Strand-Specific: | Unspecific |
| 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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