title: NextSeq 550 paired end sequencing; CYP19A1 mediated sex hormone metabolism promotes severe disease outcome in SARS-CoV-2 infected male golden hamsters
design: CYP19A1 mediated sex hormone metabolism promotes severe disease outcome in SARS-CoV-2 infected male golden hamsters
library_construction_protocol: Male and female golden hamsters (8-10 weeks old) were purchased from Janvier. All animals were kept under standard housing conditions (212C, 4050% humidity, food and water ad libitum) with a 12:12 lightdark cycle at the Leibniz Institute for Experimental Virology in Hamburg, Germany. For SARS-CoV-2 infection, hamsters were anaesthetized with 150 mg/kg ketamine and 10 mg kg-1 xylazine by intraperitoneal injection. Male and female golden hamsters (8-10 weeks old) were purchased from Janvier. All animals were kept under standard housing conditions (212C, 4050% humidity, food and water ad libitum) with a 12:12 lightdark cycle at the Leibniz Institute for Experimental Virology in Hamburg, Germany. For SARS-CoV-2 infection, hamsters were anaesthetized with 150 mg/kg ketamine and 10 mg kg-1 xylazine by intraperitoneal injection. For letrozole treatment, male and female hamsters received a final dose of 0.36 mg kg-1 d-1 or 0.18 mg kg-1 d-1 letrozole (dissolved in DMSO, Sigma-Aldrich, L6545) diluted in sterile saline (Braun), respectively, by oral gavage (stainless-steel gavaging needles) at 3 h and day 1 to 3 post infection (p.i.). From day 4 to 8 p.i., hamsters received letrozole via the drinking water as water gel packages (Necta) supplemented with 1% glucose and a final concentration of 0.0018 mg ml-1 or 0.0036 mg ml-1 letrozole for female and male hamsters, respectively. As control, hamsters were treated with DMSO as vehicle. Viral RNA was isolated from homogenized organs using the QIAamp Viral RNA Mini Kit (QIAGEN) according to the manufacturers instructions.  Preparation of stranded cDNA libraries was performed with Illumina Stranded mRNA Prep, Ligation kit, starting with 1000 ng total RNA according to the manufacturers instructions.