NASC-seq monitors RNA synthesis in single cells
2019 | journal article
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- Authors
- Hendriks, Gert-Jan; Jung, Lisa A.; Larsson, Anton J. M.; Lidschreiber, Michael; Andersson Forsman, Oscar; Lidschreiber, Katja; Cramer, Patrick ; Sandberg, Rickard
- Abstract
- Sequencing of newly synthesised RNA can monitor transcriptional dynamics with great sensitivity and high temporal resolution, but is currently restricted to populations of cells. Here, we develop new transcriptome alkylation-dependent single-cell RNA sequencing (NASC-seq), to monitor newly synthesised and pre-existing RNA simultaneously in single cells. We validate the method on pre-labelled RNA, and by demonstrating that more newly synthesised RNA was detected for genes with known high mRNA turnover. Monitoring RNA synthesis during Jurkat T-cell activation with NASC-seq reveals both rapidly up- and down-regulated genes, and that induced genes are almost exclusively detected as newly transcribed. Moreover, the newly synthesised and pre-existing transcriptomes after T-cell activation are distinct, confirming that NASC-seq simultaneously measures gene expression corresponding to two time points in single cells. Altogether, NASC-seq enables precise temporal monitoring of RNA synthesis at single-cell resolution during homoeostasis, perturbation responses and cellular differentiation.
- Issue Date
- 2019
- Journal
- Nature Communications
- ISSN
- 2041-1723
- eISSN
- 2041-1723
- Language
- English