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RNAi Reveals Phase-Specific Global Regulators of Human Somatic Cell Reprogramming
Cheng-Xu Delon Toh1, Jun-Wei Chan2, Zheng-Shan Chong2
1Epigenetics and Cell Fates Laboratory, A(∗)STAR Institute of Molecular and Cell Biology, 61 Biopolis Drive Proteos, Singapore 138673, Singapore; NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, 28 Medical Drive, Singapore 117456, Singapore.
Cell Reports
|June 14, 2016
Summary
Maximizing reprogramming efficiency requires understanding early human reprogramming mechanisms. A genome-wide RNAi screen identified key regulators, with five repressors synergistically boosting reprogramming to 85% TRA-1-60 positivity.
Area of Science:
- Cellular reprogramming
- Stem cell biology
- Genomics
Background:
- Maximizing reprogramming efficiency is crucial but hindered by incomplete mechanistic understanding.
- Identifying key regulators of early reprogramming stages is essential for improving efficiency.
Purpose of the Study:
- To conduct a systematic, genome-wide RNA interference (RNAi) screen to identify global regulators of early human reprogramming.
- To elucidate the functional roles of identified repressors and effectors in the reprogramming process.
Main Methods:
- Genome-wide RNAi screening in human cells.
- Analysis of functional repressors and effectors.
- Investigating regulatory networks in RNA processing, G protein signaling, protein ubiquitination, and chromatin modification.
- Combinatorial knockdown experiments.
Main Results:
- Identification of functional repressors and effectors impacting reprogramming.
- Discovery of interacting networks in key cellular pathways.
- Combinatorial knockdown of five repressors (SMAD3, ZMYM2, SFRS11, SAE1, ESET) achieved ~85% TRA-1-60 positivity.
- Splicing factor SFRS11 removal led to rapid pluripotency-specific splicing and regulated splicing of differentiation-related genes.
Conclusions:
- The reprogramming process involves complex, multi-layered transcriptional, splicing, and epigenetic regulation.
- SFRS11 plays a critical role in regulating alternative splicing during reprogramming.
- This study provides significant insights into the molecular machinery governing human cell reprogramming.
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