mRNA sequencing of RPE1-TP53KO and RPE1-TP53KO-EIF2AKO cells

The alternative translation initiation factor EIF2A encoded by the EIF2A gene (not to be confused with EIF2alpha, encoded by the EIF2S1 gene) plays a role in translation initiation from both conventional methionine codons, but also leucine and others upon activation of the integrated stress response. We sought to investigate the effects on global transcription of genes upon loss of EIF2A. In this experiment, EIF2A was knocked out using CRISPR-Cas9 based gene editing and confirmed using sanger sequencing and western blot in RPE1-TP53KO cells. Cells were grown to ~70% confluency and RNA was harvested using standard protocols.