Transcriptome profiling of RBM3 overexpression in A431 cells

This study aims to investigate the transcriptional changes induced by overexpression of RNA-binding protein RBM3 in A431 human epidermoid carcinoma cells. Lentiviral transduction was used to stably overexpress RBM3 in A431 cells. Total RNA was extracted from RBM3 overexpressing cells and control cells, followed by RNA-seq library preparation and sequencing on Illumina platform. Differential gene expression analysis will be performed to identify RBM3 targets and regulatory networks. A431 cells were transduced with lentivirus expressing RBM3 or EGFP control lentivirus. After puromycin selection, stable overexpressing cell lines were established and validated by western blot. Three biological replicates were prepared for each condition. Total RNA was extracted from validated stable cell lines and RNA integrity was verified. Poly(A) RNA was purified and used for strand-specific RNA-seq library construction with unique barcode indices. Libraries were multiplexed and sequenced on Illumina NovaSeq 6000 using paired-end 150bp reads.