概要: We investigated the effect of IL-13 neutralization on COVID-19 disease progression in mice.
项目整体设计: We analyzed the bulk lung tissue transcriptomes in Covid19 infected mice treated with anti-IL-13 or isotype (IgG) control antibodies and uninfected mice as an additional control group. 8-16 week-old male Tg(K18-hACE2) 2Prlmn (Jackson Laboratories) mice were challenged with 5000 plaque forming units (PFUs) of SARS-CoV-2 Hong Kong/VM20001061/2020 (BEI 14 Resources) in 50 μL by an intranasal route under 100 μL ketamine/xylazine sedation. Mice were given 150 μg of anti-IL-13(Thermofisher, clone eBio1316H; cat # 16-7135-85) or an isotype matched control IgG (Thermofisher, clone eBRG1; cat # 16-4301-85) administered on day 0, 2, and 4 pi. Mice were euthanzied at 5 days post-infection and lung tissue was resected for RNA-sequencing analysis.
Snip from lower inferior lobe was taken and RNA was extracted from murine lung tissue preserved in trizol by bead beating (tissuelyserII, Qiagen), followed by a phenol-chloroform extraction and RNA Isolation Kit (Qiagen). RNA quality was assessed using Agilent Tape station.
Ribosomal RNA was depleted using the rRNA depletion kit (NEB E6310).
建库方案:
Following rRNA 2 depletion cDNA libraries were prepared using the NEB ultra-directional library preparation kit 3 2.0 (NEB E7760) and indexed using the NEBNext Multiplex Oligos for Illumina (NEB E7335, E7500).
Libraries were diluted and pooled. 15 libraries were sequenced on the Illumina Nextseq 500 using a 150 high output kit (400 Million 8 reads, 2x75bp paired end, 150 cycles)
测序信息
分子类型:
rRNA- RNA
库的片段类型:
PAIRED
库的链类型:
Forward; -
测序平台:
ILLUMINA
测序仪型号:
Illumina NextSeq 500
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Specific; Unspecific
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文章
A specific low-density neutrophil population correlates with hypercoagulation and disease severity in hospitalized COVID-19 patients.
