RNA-seq analysis of RALD iPSCs after in vitro differentiation. RNA-seq analysis of RALD iPSCs after in vitro differentiation

We investigated roles of KRAS on stemness maintenance and differentiation propensity in the context of induced pluripotent stem cells (iPSCs) using isogenic KRAS mutant (G13C/WT) and wild-type (WT/WT) iPSCs from the same RAS-associated autoimmune lymphoproliferative syndrome-like disease (RALD) patients. RNA-seq analysis was conducted to compare gene expression profiles of WT/WT and G13C/WT iPS cells after in vitro differentiation. We found some differences of gene expression profiles regarding stemness and linage markers in the two genotypes. Overall design: To investigate the changes of stemness and linage markers between KRAS mutant and wild-type iPSCs after differentiation, the iPSCs were differentiated for 16 days . Two clones were used for each genotype (WT/WT and G13C/WT) before and after differentiation, resulting in total 8 conditions.

本研究以诱导多能干细胞（induced pluripotent stem cells, iPSCs）为模型，探究KRAS基因在干细胞干性维持及分化倾向中的调控作用，所用细胞为来自同一RAS相关自身免疫性淋巴增生综合征样疾病（RAS-associated autoimmune lymphoproliferative syndrome-like disease, RALD）患者的同基因KRAS突变型（G13C/WT）与野生型（WT/WT）诱导多能干细胞。本研究通过RNA测序（RNA-seq）分析，对比了体外分化后两类基因型细胞的基因表达谱，发现二者在干性及谱系标记基因的表达谱上存在一定差异。

整体实验设计：为探究KRAS突变型与野生型诱导多能干细胞在分化后干性及谱系标记基因的表达变化，本研究将两类细胞均体外诱导分化16天；分别收集分化前及分化后的两种基因型细胞各2个克隆，最终共得到8组实验条件。