Details

Autor(en) / Beteiligte

• 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

Titel

RNAi Reveals Phase-Specific Global Regulators of Human Somatic Cell Reprogramming

Ist Teil von

• Cell reports (Cambridge), 2016-06, Vol.15 (12), p.2597-2607

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• 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. [Display omitted] •Key functional regulators of human somatic cell reprogramming are identified•Combinatorial knockdown of five repressors allows highly efficient reprogramming (85%)•SFRS11 regulates splicing of genes that are critical for reprogramming•Splicing of downstream ZNF207 isoform B to A and C affects reprogramming efficiency Toh et al. identified genome-wide functional regulators of the early stages of human somatic cell reprogramming in a phase-specific RNAi screen. Combinatorial depletion of the top repressors allows reprogramming to proceed unhindered at a near-deterministic efficiency. SFRS11, an mRNA splicer, blocks reprogramming partially through the splicing of ZNF207 isoforms.