An Unbiased Approach to Identifying Cellular Reprogramming-Inducible Enhancers.
Details
Serval ID
serval:BIB_3AC9363F86AE
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
An Unbiased Approach to Identifying Cellular Reprogramming-Inducible Enhancers.
Journal
International journal of molecular sciences
ISSN
1422-0067 (Electronic)
ISSN-L
1422-0067
Publication state
Published
Issued date
06/12/2024
Peer-reviewed
Oui
Volume
25
Number
23
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Publication Status: epublish
Abstract
Cellular reprogramming of somatic cells towards induced pluripotency is a multistep stochastic process mediated by the transcription factors Oct4, Sox2, Klf4 and c-Myc (OSKM), which orchestrate global epigenetic and transcriptional changes. We performed a large-scale analysis of integrated ChIP-seq, ATAC-seq and RNA-seq data and revealed the spatiotemporal highly dynamic pattern of OSKM DNA binding during reprogramming. We found that OSKM show distinct temporal patterns of binding to different classes of pluripotency-related enhancers. Genes involved in reprogramming are regulated by the coordinated activity of multiple enhancers, which are sequentially bound by OSKM for strict transcriptional control. Based on these findings, we developed an unbiased approach to identify Reprogramming-Inducible Enhancers (RIEs), constructed enhancer-traps and isolated cells undergoing reprogramming in real time. We used a representative RIE taken from the Upp1 gene fused to Gfp and isolated cells at different time-points during reprogramming and found that they have unique developmental capacities as they are reprogrammed with high efficiency due to their distinct molecular signatures. In conclusion, our experiments have led to the development of an unbiased method to identify and isolate reprogrammable cells in real time by exploiting the functional dynamics of OSKM, which can be used as efficient reprogramming biomarkers.
Keywords
Cellular Reprogramming/genetics, Kruppel-Like Factor 4/metabolism, Animals, Enhancer Elements, Genetic, Mice, Octamer Transcription Factor-3/genetics, Octamer Transcription Factor-3/metabolism, Induced Pluripotent Stem Cells/metabolism, Induced Pluripotent Stem Cells/cytology, SOXB1 Transcription Factors/genetics, SOXB1 Transcription Factors/metabolism, Kruppel-Like Transcription Factors/genetics, Kruppel-Like Transcription Factors/metabolism, Transcription Factors/metabolism, Transcription Factors/genetics, Proto-Oncogene Proteins c-myc/metabolism, Proto-Oncogene Proteins c-myc/genetics, Chromatin Immunoprecipitation Sequencing/methods, OSKM, cellular reprogramming, chromatin structure, enhancers, genomics, iPSCs, transcription factors, transcriptional regulation
Pubmed
Open Access
Yes
Create date
20/12/2024 12:04
Last modification date
21/12/2024 7:09