RRC ID 72681
Author Kouno T, Moody J, Kwon AT, Shibayama Y, Kato S, Huang Y, Böttcher M, Motakis E, Mendez M, Severin J, Luginbühl J, Abugessaisa I, Hasegawa A, Takizawa S, Arakawa T, Furuno M, Ramalingam N, West J, Suzuki H, Kasukawa T, Lassmann T, Hon CC, Arner E, Carninci P, Plessy C, Shin JW.
Title C1 CAGE detects transcription start sites and enhancer activity at single-cell resolution.
Journal Nat Commun
Abstract Single-cell transcriptomic profiling is a powerful tool to explore cellular heterogeneity. However, most of these methods focus on the 3'-end of polyadenylated transcripts and provide only a partial view of the transcriptome. We introduce C1 CAGE, a method for the detection of transcript 5'-ends with an original sample multiplexing strategy in the C1TM microfluidic system. We first quantifiy the performance of C1 CAGE and find it as accurate and sensitive as other methods in the C1 system. We then use it to profile promoter and enhancer activities in the cellular response to TGF-β of lung cancer cells and discover subpopulations of cells differing in their response. We also describe enhancer RNA dynamics revealing transcriptional bursts in subsets of cells with transcripts arising from either strand in a mutually exclusive manner, validated using single molecule fluorescence in situ hybridization.
Volume 10(1)
Pages 360
Published 2019-1-21
DOI 10.1038/s41467-018-08126-5
PII 10.1038/s41467-018-08126-5
PMID 30664627
PMC PMC6341120
MeSH A549 Cells Animals Cell Line Enhancer Elements, Genetic* Fibroblasts / cytology Fibroblasts / drug effects Fibroblasts / metabolism* Gene Expression Profiling Humans In Situ Hybridization, Fluorescence Mice Microfluidic Analytical Techniques Promoter Regions, Genetic RNA, Messenger / genetics* RNA, Messenger / metabolism Sequence Analysis, RNA Single-Cell Analysis / instrumentation Single-Cell Analysis / methods* Transcription Initiation Site* Transcriptome* Transforming Growth Factor beta / pharmacology
IF 12.121
Resource
Human and Animal Cells B6G-2(AES0003) iPS-MEF-Ng-20D-17(APS0001)