Mutation calls by SMM-Seq of MutaMouse male liver exposed to Benzo(B)Fluoranthene for 28 days
Description
MutaMouse males were exposed to five doses of Benzo(B)Fluoranthene (BbF) alongside vehicle controls by oral gavage for 28 days with a 3 day sampling time. Random whole-genome sequencing of liver DNA was performed using MutagenTech's SMM-Seq. Each sample was sequenced to roughly 200Mb. Data is single-nucleotide variants identified by SMM-Seq for 24 mouse liver samples. Variants identified by SMM-seq were contrasted against the single-nucleotide polymorphisms database (dbSNP) to remove known mouse variants. Additionally, germline variants were excluded through conventional whole-genome sequencing of matched samples at 20X depth. Mutation frequency (number of mutations divided by total read depth) is significantly increased by BbF at the top two doses compared to control. BbF increases proportions of C>A subtypes compared to control.
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Steps to reproduce
MutaMouse animals (CD2F1 BALB/c X DBA/2 strain) 8-10 weeks old were exposed to 0, 6.25, 12.5, 25, 50, or 100 mg/kg body weight/day Benzo[B]Fluoranthene (BbF) dissolved in olive oil for 28 consecutive days by oral gavage. Liver tissue was harvested from euthanized animals 3 days after the end of exposure and flash frozen at -80C until analysis. DNA was extracted using the Qiagen DNeasy blood and tissue kit protocol (Cat. #69504, Qiagen, Hilden, Germany) with modifications: tissue disruption by bead beating samples twice at 4000 rpm for 30s and proteinases K digestion was conducted at 37C for 180min. Libraries were prepared and sequenced as described in Maslov et al., 2022 (10.1126/sciadv.abm3259).