RNA-seq in SREBF1-deficient human iPSCs, hiPSC-NSCs and differentiated cells.

Omics analyses and qRT-PCR time-course during the transition from hiPSC to hiPSC-NSC highlighted the up-regulation of SREBF1, a gene involved in cholesterol biosynthesis and lipid homeostasis, suggesting its potential role in NSC commitment/maintenance. To test this hypothesis, we generated SREBF1-deficient hiPSC lines by co-delivering ribonucleoprotein Cas9 with a pool of sgRNA targeting exon 5 of the SREBF1 gene. Upon isolation of three clones harboring the deletion we performed RNA-seq analysis in hiPSC, hiPSC-NSC and differentiated cultures at 7 and 14 days of differentiation, compared to control cells. This analysis will allow to identify the potential role of SREBF1 in affecting hiPSC-to-hiPSC-NSC transition, hiPSC-NSC maintenance and commitment toward differentiated cell populations. Overall design: We performed the poly-A+ RNA-seq profiling in three clones of SREBF1-deficient human induced pluripotent stem cells (hiPSCs), neural stem cells (hiPSC-NSCs) and differentiated cells at d7 and d14. We also included three replicates of negative control for each differentiation stage.