Transcriptome analysis (bulk RNAseq) of FlncKO-C2C12 during myogenic differentiation
In this study we performed transcriptome analysis (bulk RNAseq) of WT-C2C12 immortalized mouse myoblasts and FlncKO-C2C12 cells during myogenic differentiation (day 0, 1, 2, 4 and 6). Overall design: C2C12 cells were genetically modified using CRISPR/Cas9 guide RNA targeting the first exon of the mouse Flnc gene (NC_000072.6 (2933153.29461889)) as described in Klimenko et al., 2024. WT-C2C12 (wild type)and FlncKO-C2C12 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM, 4.5 g/L d-Glucose, Gibco) supplemented with 10 % fetal bovine serum (FBS, Gibco), 2 mM l-glutamine, and 100 µg/ml penicillin-streptomycin (Pen-Strep, Gibco). Cells were re-plated every 48 h at a density of 100,000 cells per 100 mm dish. Myogenic differentiation was induced by culturing cells in low serum medium (DMEM, 2 mM l-glutamine, 100 µg/ml Pen-Strep) supplemented with 2 % horse serum (Gibco). During differentiation, the medium was refreshed daily. Total RNA was extracted on 0, 1, 2, 4 and 6 days after induction of myogenic differentiation. Total RNA was extracted using ExtractRNA reagent (Evrogen) according to the manufacturer's instructions. RNA-libraries from WT and FlncKO cells were prepared with TruSeq Stranded mRNA kit, samples were sequenced on Illumina NextSeq 2000, read length 51 bp.