RNA Sequencing Facilitates Quantitative Analysis of Transcriptomes of human early hematopoietic differentiation

Purpose: The goals of this study are to identify key transcription factors governing differentiation through comparing thel transcriptome profilings in hESC samples collected from day 0 to day 4 after hematopoietic differentiation. Conclusions: 68 transcriptional factors were found up-regulated gradually and steadily during early hematopoietic differentiation of H1 hESCs. After 4 days of differentiation, the mRNA levels of all these factors increased by more than 10 folds. Overall design: Methods: mRNA profiles of hESC samples collected from day 0 to day 4 after hematopoiesis differentiation were generated by deep sequencing. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays