Genome-wide Functional Analysis Reveals Factors Needed at the Transition Steps of Induced Reprogramming

•A genome-wide RNAi screen identified genes required for various reprogramming steps•Non-differentially expressed genes play important roles in cell-fate transitions•Dmbx1, Hnf4g, Nobox, and Asb4 are essential for efficient reprogramming•Nfe2, Cdkn2aip, Msx3, Dbx1, Lzts1, Arx, Gtf2i, and Ankrd22 compromise reprogramming

SummaryAlthough transcriptome analysis can uncover the molecular changes that occur during induced reprogramming, the functional requirements for a given factor during stepwise cell-fate transitions are left unclear. Here, we used a genome-wide RNAi screen and performed integrated transcriptome analysis to identify key genes and cellular events required at the transition steps in reprogramming. Genes associated with cell signaling pathways (e.g., Itpr1, Itpr2, and Pdia3) constitute the major regulatory networks before cells acquire pluripotency. Activation of a specific gene set (e.g., Utf1 or Tdgf1) is important for mature induced pluripotent stem cell formation. Strikingly, a major proportion of RNAi targets (∼53% to 70%) includes genes whose expression levels are unchanged during reprogramming. Among these non-differentially expressed genes, Dmbx1, Hnf4g, Nobox, and Asb4 are important, whereas Nfe2, Cdkn2aip, Msx3, Dbx1, Lzts1, Gtf2i, and Ankrd22 are roadblocks to reprogramming. Together, our results provide a wealth of information about gene functions required at transition steps during reprogramming.

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