RNA-seq of 293T cells expressing different RUNX1 splice isoforms

Biological relevance: RUNX1b is a transcription factor that is heavily implicated in leukemogenesis. In this study we identified that the RNA-binding protein hnRNP K binds to the RUNX1b transcript resulting in an alternatively spliced transcript that lacks Exon4. To determine the transcriptional consequences of this alternatively spliced variant, we performed RNA-Seq. Intent: To determine the effect of overexpression of RUNX1b or RUNX1b dEx4 Experimental Workflow: 293T cells were transduced with either empty vector, plasmids expressing RUNX1b full-length isoform or the RUNX1b dEx4 isoform. All plasmids used were doxycycline inducible. The cells were induced with doxycycline (0.4 mg/ml) for 72 hours, following which RNA was collected and sequenced. All experiments were done in triplicate.