RNA Sequencing Facilitates Quantitative Analysis of Wild Type and MEIS2 deleted H1 drived cells at day4 of hematopoietic differentiation.

Purpose: The goals of this study are to investigate the molecular mechanism by which MEIS2 controls HEP specification and EHT through compareing the mRNA profiling of Wild Type and MEIS2 deleted H1 drived cells at day4 of hematopoietic differentiation. Methods: mRNA profiles of Wild Type and MEIS2 deleted H1 drived cells at day4 of hematopoietic differentiation were generated by deep sequencing using Illumina GAIIx. 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 Conclusions:a large number of genes were down-regulated in MEIS2-deleted H1 hESCs when compared with the wild-type cells. Among those, a number of mammalian hematopoiesis-associated genes such as TAL1 ,GFI and GATA2 were significantly down-regulated. mRNA profiles of Wild Type and MEIS2 deleted H1 drived cells at day4 of hematopoietic differentiation were generated by deep sequencing using Illumina GAIIx.