Related Experiment Video
Updated: Jun 13, 2025
Cell Surface Marker Mediated Purification of iPS Cell Intermediates from a Reprogrammable Mouse Model
Published on: September 6, 2014
Single-cell profiling identifies LIN28A mRNA targets in the mouse pluripotent-to-2C-like transition and somatic cell
Jieyi Hu1, Jianwen Yuan2, Quan Shi3
1Laboratory of Integrative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China; University of Chinese Academy of Sciences, Beijing, China.
Abstract:
RNA-binding proteins (RBPs) regulate totipotency, pluripotency maintenance, and induction. The intricacies of how they modulate these processes through their interaction with RNAs remain to be elucidated. Here we employed Targets of RBPs Identified By Editing (TRIBE) with single-cell resolution (scTRIBE) to profile the mRNA targets of the key pluripotency regulator LIN28A in mouse embryonic stem cells (ESCs), 2-cell embryo-like cells (2CLCs), and somatic cell reprogramming. LIN28A is known to act by controlling the maturation of the let-7 microRNA, but, in addition, it binds to multiple mRNAs and influences their stability and translation efficiency. However, the mRNA targets of LIN28A in 2CLCs and reprogramming are unclear. Through quantitative single-cell analysis of the scTRIBE dataset, we observed a marked increase in the binding of LIN28A to mRNAs of ribosome biogenesis factors and a selected group of totipotency factors in 2CLCs within ESC cultures. Our results suggest that LIN28A extends the half-life of at least some of these mRNAs, providing new insights into its role in the totipotent state. We also uncovered the distinct trajectory-specific LIN28A-mRNA networks in reprogramming, helping explain how LIN28A facilitates the mesenchymal-to-epithelial transition and pluripotency acquisition. Our study not only clarifies the multifunctional role of LIN28A in these processes but also highlights the importance of decoding RNA-protein interactions at the single-cell level.
Insights
RNA-binding proteins like LIN28A are crucial for cell identity. This study reveals LIN28A
Area of Science:
- Molecular Biology
- Stem Cell Biology
- Epigenetics
Background:
- RNA-binding proteins (RBPs) are critical regulators of cellular processes, including stem cell pluripotency and development.
- LIN28A is a key pluripotency regulator known to control microRNA maturation and mRNA translation/stability, but its mRNA targets in specific cell states remain incompletely understood.
- Understanding RBP-RNA interactions at a single-cell level is essential for deciphering complex biological processes like cell reprogramming and totipotency.
Purpose of the Study:
- To profile the mRNA targets of the RNA-binding protein LIN28A in mouse embryonic stem cells (ESCs), 2-cell embryo-like cells (2CLCs), and during somatic cell reprogramming.
- To elucidate the role of LIN28A in maintaining totipotency and facilitating pluripotency acquisition during cellular reprogramming.
- To investigate LIN28A-mediated RNA regulation at single-cell resolution.
Main Methods:
- Application of Targets of RBPs Identified By Editing (TRIBE) with single-cell resolution (scTRIBE) to map LIN28A-bound mRNAs.
- Quantitative single-cell analysis of scTRIBE datasets.
- Analysis of LIN28A-mRNA networks during somatic cell reprogramming.
Main Results:
- A significant increase in LIN28A binding to mRNAs encoding ribosome biogenesis factors and totipotency factors was observed in 2CLCs.
- LIN28A was found to extend the half-life of specific totipotency-related mRNAs, suggesting a role in maintaining the totipotent state.
- Distinct LIN28A-mRNA networks were identified in specific reprogramming trajectories, correlating with mesenchymal-to-epithelial transition and pluripotency acquisition.
Conclusions:
- LIN28A plays a multifaceted role in regulating stem cell states, extending beyond its known function in microRNA processing.
- LIN28A's interaction with specific mRNAs is crucial for maintaining totipotency and facilitating cellular reprogramming.
- Single-cell resolution of RNA-protein interactions provides critical insights into the dynamic regulation of gene expression in stem cells and reprogramming.
More Related Videos
Related Concept Videos
01:29Somatic to iPS Cell Reprogramming
01:24Methods of Nuclear Reprogramming
01:32Chromatin Modification in iPS Cells
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
01:21Lineage Commitment