Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

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RNA-Seq of UPF1 depletion in human colorectal adenocarcinoma cell line HCT116 or human embryonic kidney cell line HEK293 via the dTAG degron system


ABSTRACT: UPF1 is a multi-domain RNA helicase that constantly monitors the transcriptome by non-specifically binding to mRNAs, dissociating from non-target transcripts, and initiating degradation on selected target RNAs via multiple proposed pathways such as nonsense-mediated decay (NMD). NMD is a translation-coupled mechanism that targets mRNAs harboring a premature stop codon (PTC) for degradation, thereby serving as a quality control and gene regulatory pathway ensuring transcriptome integrity. The UPF1 gene is essential in cultured human cells and previous studies relied mostly on RNA interference to downregulate UPF1. Here we established an dTAGV-1-inducible UPF1 degron system in the human colorectal adenocarcinoma cell line HCT116 or human embryonic kidney cell line HEK293 by tagging UPF1 at the N-terminus with an Myc-FKBP-tag (FKBP = FKBP12-F36V). With these cell lines we wanted to explore the transcriptome-wide expression changes including the extent of NMD inhibition upon rapid depletion of UPF1. To this end, depletion of UPF1 was induced with 0.25 µM dTAGV-1 for 12h. As controls, the parental cell line or untreated cells were used.

INSTRUMENT(S): Illumina NovaSeq 6000

ORGANISM(S): Homo sapiens

SUBMITTER: Volker Böhm 

PROVIDER: E-MTAB-13829 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications


The RNA helicase UPF1 shapes the transcriptome as the core factor of nonsense-mediated mRNA decay (NMD). The essential role of UPF1 in human cells has impeded efforts to delineate its directly regulated transcripts and molecular function. To investigate the effects of rapid UPF1 depletion, we engineered human cell lines with endogenous UPF1 fused to conditional degron tags. Temporal-resolution transcriptomic analyses identified direct target mRNAs, consisting predominantly of NMD substrates that  ...[more]

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