RNA-seq of mouse wild-type and I830077J02Rik,C130074G19Rik,1500009L16Rik,D630003M21Rik,D430041D05Rik and A530016L24Rik gene knockouts in Flk-1+/Pdgfra+ and Flk-1+/Pdgfra- sorted populations at day 4 of embryoid body differentiation
Haematopoietic and endothelial cells derive from a subset of Flk-1 expressing cells. In vitro differentiation of mouse embryonic stem cells (mESCs) to embryoid bodies (EBs) recapitulate several aspects of early embryonic development, including the formation of a subset of Flk-1 expressing cells. By day 4 of EB differentiation, mesodermal lineages with distinct lineage potentials can be identified based on the expression of the two surface markers Flk-1 and Pdgfra. Cells expressing Flk-1 alone (Flk-1?/Pdgfra?) are enriched for haematoendothelial precursors, whereas double-positive cells (Flk-1?/Pdgfra?) are enriched for primitive/cardiac mesodermal lineages.  We previously identified six uncharacterized genes (Riken) as potential regulators of haematoendothelial or cardiac lineages commitment, namely I830077J02Rik,C130074G19Rik,1500009L16Rik,D630003M21Rik,D430041D05Rik and A530016L24Rik. We generated homozygous knockouts (KOs) of each of these 6 genes and performed bulk RNA sequencing of WT cells and gene KOs is the two sorted populations (Flk-1?/Pdgfra? and Flk-1?/Pdgfra+) at day 4 of embryoid body differentiation.