RNAi Reveals Phase-Specific Global Regulators of Human Somatic Cell Reprogramming
Author(s)Toh, Cheng-Xu Delon; Chan, Jun-Wei; Chong, Zheng-Shan; Wang, Hao Fei; Guo, Hong Chao; Satapathy, Sandeep; Ma, Dongrui; Goh, Germaine Yen Lin; Khattar, Ekta; Yang, Lin; Tergaonkar, Vinay; Chang, Young-Tae; Collins, James J.; Daley, George Q.; Wee, Keng Boon; Farran, Chadi A. EL; Li, Hu; Lim, Yoon-Pin; Bard, Frederic A.; Loh, Yuin-Han; Collins, James J.; ... Show more Show less
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Incomplete knowledge of the mechanisms at work continues to hamper efforts to maximize reprogramming efficiency. Here, we present a systematic genome-wide RNAi screen to determine the global regulators during the early stages of human reprogramming. Our screen identifies functional repressors and effectors that act to impede or promote the reprogramming process. Repressors and effectors form close interacting networks in pathways, including RNA processing, G protein signaling, protein ubiquitination, and chromatin modification. Combinatorial knockdown of five repressors (SMAD3, ZMYM2, SFRS11, SAE1, and ESET) synergistically resulted in ∼85% TRA-1-60-positive cells. Removal of the novel splicing factor SFRS11 during reprogramming is accompanied by rapid acquisition of pluripotency-specific spliced forms. Mechanistically, SFRS11 regulates exon skipping and mutually exclusive splicing of transcripts in genes involved in cell differentiation, mRNA splicing, and chromatin modification. Our study provides insights into the reprogramming process, which comprises comprehensive and multi-layered transcriptional, splicing, and epigenetic machineries.
DepartmentInstitute for Medical Engineering and Science; MIT Synthetic Biology Center; Massachusetts Institute of Technology. Department of Biological Engineering
Toh, Cheng-Xu Delon, Jun-Wei Chan, Zheng-Shan Chong, Hao Fei Wang, Hong Chao Guo, Sandeep Satapathy, Dongrui Ma, et al. "RNAi Reveals Phase-Specific Global Regulators of Human Somatic Cell Reprogramming." Cell Reports 15:12 (21 June 2016), pp.2597-2607.
Final published version