title: RNA-seq of human primary epithelial nasal cells infected with SARS-CoV-2 (20A clade) and treated with long chain inorganic polyphosphates (polyP120) against vehicle-(saline solution)- treated controls
design: RNA-seq of human primary epithelial nasal cells infected with SARS-CoV-2 (20A clade) and treated with long chain inorganic polyphosphates (polyP120) against vehicle-(saline solution)- treated controls
library_construction_protocol: Primary human epithelial cells from nasal brushing were plated (3x105 cells) and infected with SARS-CoV-2 viral particles 20A (EPI_ISL_514432-S66; MOI, 0.03). After 24 hours, the infected cells were treated with 18.5 microMolar long chain inorganic polyphosphates (polyP120) or vehicle (saline solution) as the negative control for the treatment. After 36 hours, the cells were lysed with TRIzol RNA Isolation Reagent (#15596018; Ambion, Thermo Fisher Scientific) and frozen at -80°C. Freshly isolated human nasal epithelial cells were collected by nasal brushing of healthy donors (data deposited at: EVA- EMBL-EBI; project ID: PRJEB42411; analyses: ERZ1700617). These cells were cultured in PneumaCult (#05009; StemCell Technologies) with 2 mM L-glutamine (25030-024; Gibco), and 1% penicillin/streptomycin (P0781; Sigma).  Primary human epithelial cells from nasal brushing were plated (300000 cells) and infected with SARS-CoV-2 viral particles 20A (EPI_ISL_514432-S66; MOI, 0.03). After 24 hours, the infected cells were treated with 18.5 microMolar long chain inorganic polyphosphates (polyP120) or vehicle (saline solution) as the negative control for the treatment. After 36 hours, the cells were lysed, and their RNA were extracted. Total RNA, isolated from human primary nasal epithelial cells by using TRIzol RNA Isolation Reagent (#15596018; Ambion, Thermo Fisher Scientific) was quantified in a NanoDrop™ One/OneC Microvolume UV-Vis Spectrophotometer (Thermo Scientific™), checked for purity and integrity.  Libraries were prepared using the TruSeq Stranded Total RNA LT Sample Prep Kit (Gold) according to the protocols recommended by the manufacturer. Library kit: TruSeq Stranded Total RNA LT Sample Prep Kit (Gold); Library protocol: TruSeq Stranded Total RNA Sample Prep Guide, Part # 15031048 Rev. E; Reagent: NovaSeq 6000 S4 Reagent Kit;