Genome Wide Binding profile and transcriptome of STAT3 and STAT3 Mutant Y640F in STAT3 Wildtype and STAT3 Mutant Y640F hematopoietic progenitor cell lines

In many human cancers the transcription factor STAT3 is constitutively active and contributes to oncogenesis, tumour growth and progression. Recently, gain-of-function mutations of STAT3 have been identified in patients suffering from various haematopoietic malignancies and are postulated to enhance the transcriptional activity of STAT3. We investigated the gene expression and binding profile of the most common STAT3 mutant Y640F compared to wild-type STAT3, describing it's mechanism of action and pinpointing to novel therapeutic intervention sites. (RNA-Seq) mRNA from HPC7 or HPCLSK transduced with retroviral vectors encoding empty vector, STAT3 or STAT3Y640F (3 technical replicates) were prepared using the Lexogen SENSE mRNA-Seq library preparation kit. Single-end, 50bp sequencing was performed on an Illumina HiSeq 3000/4000 sequencer. Reads were mapped to the GENECODE M13 genome. (ChIP Seq) HPC7 expressing empty vector, STAT3, STAT3Y640F ) linked to a C-terminal V5 tag were used for V5-ChIP (3 technical replicates). After V5-pull down genomic regions were subjected to sequencing library preparation and sequencing was performed using the Illumina HiSeq3000/4000 platform in 50-base-pair single-end mode.