NextSeq 500 paired end sequencing; RNA-seq of mouse Embryonic Stem Cell (mESC) line-derived endothelial and hematopoietic cells subjected to differential doxycycline (dox)-induced expression of transcription factors (TFs) in a Tal1Δ/Δ genetic background
RNA-seq of mouse Embryonic Stem Cell (mESC) line-derived endothelial and hematopoietic cells subjected to differential doxycycline (dox)-induced expression of transcription factors (TFs) in a Tal1Δ/Δ genetic background
The doxycycline-inducible ESC line was generated using the inducible cassette exchange method, the constructs and plasmids described in Bergiers et al., 2018; Iacovino et al., 2011; Vargel et al., 2016. Through gene synthesis (GenScript) and cloning, the p2lox-3TFs plasmids was generated and consequently transfected into A2.lox ESCs (kindly provided by Dr. Michael Kyba – University of Minnesota). The deletion of the Tal1 gene to generate the i3TFs Tal1Δ/Δ mESC lines was achieved using the Crispr/CAS9 technology as described in Serina Secanechia et al., 2022. ESCs were maintained, expanded and differentiated as described in Serina Secanechia et al., 2022. Cells were FACS sorted from the hematopoietic 'hemangioblast culture' as described in Serina Secanechia et al., 2022. RNA-seq library preparation and sequencing were performed as described in Picelli et al., 2014 and Serina Secanechia et al., 2022