RNA sequencing of Sphk1fl/fl Sphk2-/- and Sphk1-/- Sphk2-/- mESC clones.

We report a growth defect in sphingosine kinase null clones due to a cell cycle arrest in G2/M phase resulting in a growth defect. GSEA showed upregulation of the G2M checkpoint geneset. Further, telomere elongation factors are upregulated in kinase null cells. Overall design: mESC clones derived from Sphk1fl/fl Sphk2-/- mice were cultured in tamoxifen for 48 hours then sorted for Cre-GFP expression yielding GFP- Sphk1fl/fl Sphk2-/- cells and GFP+ Sphk1-/- Sphk2-/- (Kinase Null) cells. This was done for 2 separate clones resulting in 4 sample types: Clone 4 GFP-, Clone 4 GFP+, Clone 10 GFP- and Clone 10 GFP+.

本研究报道了鞘氨醇激酶（sphingosine kinase）敲除克隆的生长缺陷，该缺陷由G2/M期细胞周期阻滞所引发。基因集富集分析（Gene Set Enrichment Analysis, GSEA）结果显示，G2M检查点基因集呈现上调表达。此外，在激酶敲除细胞中，端粒延长相关因子呈现上调表达。总体实验设计：本研究使用源自Sphk1fl/fl Sphk2-/-小鼠的小鼠胚胎干细胞（mouse embryonic stem cell, mESC）克隆，将其在他莫昔芬中培养48小时后，基于Cre-绿色荧光蛋白（Cre-GFP）的表达进行分选，得到GFP阴性的Sphk1fl/fl Sphk2-/-细胞与GFP阳性的Sphk1-/- Sphk2-/-（激酶敲除）细胞。该实验针对2个独立克隆开展，最终得到4种样本类型：克隆4 GFP阴性组、克隆4 GFP阳性组、克隆10 GFP阴性组以及克隆10 GFP阳性组。