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| Item Type: | Article |
|---|---|
| Title: | SLAM-Drop-seq reveals mRNA kinetic rates throughout the cell cycle |
| Creators: |
Liu, H.  ORCID: https://orcid.org/0009-0007-4871-452X, Arsiè, R. ORCID: https://orcid.org/0000-0002-7869-7624, Schwabe, D. ORCID: https://orcid.org/0000-0003-2825-3352, Schilling, M. ORCID: https://orcid.org/0000-0002-3453-7792, Minia, I. ORCID: https://orcid.org/0000-0003-1012-3373, Alles, J. ORCID: https://orcid.org/0000-0003-0681-0410, Boltengagen, A. ORCID: https://orcid.org/0000-0002-8356-9766, Kocks, C. ORCID: https://orcid.org/0000-0003-1749-3334, Falcke, M. ORCID: https://orcid.org/0000-0001-7137-1114, Friedman, N. ORCID: https://orcid.org/0000-0002-9678-3550, Landthaler, M. ORCID: https://orcid.org/0000-0002-1075-8734 and Rajewsky, N. ORCID: https://orcid.org/0000-0002-4785-4332
|
| Abstract: | RNA abundance is tightly regulated in eukaryotic cells by modulating the kinetic rates of RNA production, processing, and degradation. To date, little is known about time-dependent kinetic rates during dynamic processes. Here, we present SLAM-Drop-seq, a method that combines RNA metabolic labeling and alkylation of modified nucleotides in methanol-fixed cells with droplet-based sequencing to detect newly synthesized and preexisting mRNAs in single cells. As a first application, we sequenced 7280 HEK293 cells and calculated gene-specific kinetic rates during the cell cycle using the novel package Eskrate. Of the 377 robust-cycling genes that we identified, only a minor fraction is regulated solely by either dynamic transcription or degradation (6 and 4%, respectively). By contrast, the vast majority (89%) exhibit dynamically regulated transcription and degradation rates during the cell cycle. Our study thus shows that temporally regulated mRNA degradation is fundamental for the correct expression of a majority of cycling genes. SLAM-Drop-seq, combined with Eskrate, is a powerful approach to understanding the underlying mRNA kinetics of single-cell gene expression dynamics in continuous biological processes. |
| Keywords: | Cell Cycle, mRNA Kinetics, Single Cells, Temporal Regulation, Transcription and Degradation |
| Source: | Molecular Systems Biology |
| ISSN: | 1744-4292 |
| Publisher: | Wiley |
| Volume: | 19 |
| Number: | 10 |
| Page Range: | e11427 |
| Date: | 12 October 2023 |
| Official Publication: | https://doi.org/10.15252/msb.202211427 |
| PubMed: | View item in PubMed |
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