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Endogenous mRNA decay in 293t, hela, RPE cell line

ABSTRACT: endogenous mRNA decay in 293t, hela, RPE cell line

PROVIDER: PRJNA522261 | ENA |

REPOSITORIES: ENA

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endogenous mRNA decay in 293t, hela, RPE cell line

Project description:mRNA endogenous decay profiles approach in 293T, HeLa and RPE cells. mRNA translation decodes nucleotide into amino acid sequences. However, translation has also been shown to affect mRNA stability depending on codon composition in model organisms, although universality of this mechanism remains unclear. Here, using three independent approaches to measure exogenous and endogenous mRNA decay, we define which codons are associated with stable or unstable mRNAs in human cells. We demonstrate that the regulatory information affecting mRNA stability is encoded in codons and not in nucleotides. Stabilizing codons tend to be associated with higher tRNA levels and higher charged/total tRNA ratios. While mRNAs enriched in destabilizing codons tend to possess shorter poly(A)-tails, the poly(A)-tail is not required for the codon-mediated mRNA stability. This mechanism depends on translation; however, the number of ribosome loads into a mRNA modulates the codon-mediated effects on gene expression. This work provides definitive evidence that translation strongly affects mRNA stability in a codon-dependent manner in human cells.

2019-05-01 | GSE126520 | GEO

ORFome decay in 293t and k562

Project description:ORFome decay in 293t and k562

| PRJNA522263 | ENA

ORFome decay in 293t and k562

Project description:mRNA translation decodes nucleotide into amino acid sequences. However, translation has also been shown to affect mRNA stability depending on codon composition in model organisms, although universality of this mechanism remains unclear. Here, using three independent approaches to measure exogenous and endogenous mRNA decay, we define which codons are associated with stable or unstable mRNAs in human cells. We demonstrate that the regulatory information affecting mRNA stability is encoded in codons and not in nucleotides. Stabilizing codons tend to be associated with higher tRNA levels and higher charged/total tRNA ratios. While mRNAs enriched in destabilizing codons tend to possess shorter poly(A)-tails, the poly(A)-tail is not required for the codon-mediated mRNA stability. This mechanism depends on translation; however, the number of ribosome loads into a mRNA modulates the codon-mediated effects on gene expression. This work provides definitive evidence that translation strongly affects mRNA stability in a codon-dependent manner in human cells. 293T cells and k562 cells were infected with ORFome library to decay exodogenous gene mRNA decay. Stable infected cells were treated with actinmycin D at 6 well plate and samples were collected in duplicate every hour 0-6h for RNA-seq

2019-05-01 | GSE126521 | GEO

Mitochondrial RNA-seq and total RNA-seq of 293T, HeLa, and N2a cells; RIP-seq of SUPV3L1, ceSUPV3L1, ATP5B, and TRAP1; Small RNA-seq of 293T mitochondrial fractions

Project description:The mechanism of mecciRNA degradation remains unknown. To investigate the degradation of mecciRNAs, we performed mitochondrial RNA sequencing and total RNA sequencing of 293T, HeLa, and N2a cells. To investigate the degradation mechanism of mecciRNAs, RIP-seq was conducted in 293T and C. elegans. Small RNA sequencing of mitochondrial sucrose gradient fractions was performed to identify mecciRNA degradation fragments.

2023-12-19 | GSE249786 | GEO

SLAM-seq for K562 endogenous mRNA decay

Project description:SLAM-seq for K562 endogenous mRNA decay

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RNA-seq of KD, rescues of NMD factors, and UPF1-flag CLIP-seq in HeLa cells.

Project description:This study aims at confidently identifying endogenous nonsense mediated decay (NMD) targets. To achieve this purpose, we performed KD of a few NMD factors in HeLa cells. Additionally, we performed rescue experiments for each factor, expressing an RNAi-resistant version of the gene from a plasmid. To determine transcripts bound by UPF1 in HeLa cells, A construct with a C-terminally flag tagged version of UPF1 was expressed. In order to avoid competition with endogenous UPF1, a KD was performed.

2016-12-19 | GSE86148 | GEO

Genome-wide analysis of Human Upf1, SMG5, SMG7 and PNRC2 downregulated substrates in HeLa cell

2012-11-01 | GSE34876 | GEO

Genome-wide analysis of Human Upf1, SMG5, SMG7 and PNRC2 downregulated substrates in HeLa cell

Project description:Analysis of cellular NMD (Nonsense-mediated mRNA decay) substrates that regulated by Upf1, SMG5, SMG7 and/or PNRC2 in HeLa cell. The hypothesis tested in the present study was that endogenous NMD substrates may co-regulated by Upf1, SMG5, SMG7 and PNRC2. Total RNA obtained from HeLa cells with downregulation of Upf1, SMG5, PNRC2 or SMG7 by siRNA. The up- or down-regulated transcripts were compare to control siRNA treated HeLa cell RNA extract. Significant transcripts were confirmed by replication

2012-11-01 | E-GEOD-34876 | biostudies-arrayexpress

Translation is required for miRNA-dependent decay of endogenous transcripts.

Project description:Translation is required for miRNA-dependent decay of endogenous transcripts.

| PRJNA599577 | ENA

Genome-wide analysis of cellular SMD or NMD substrates that regulated by Upf1 or PNRC2 in HeLa cell

Project description:Analysis of cellular SMD or NMD substrates that regulated by Upf1 and/or PNRC2 in HeLa cell. The hypothesis tested in the present study was that endogenous SMD or NMD substrates may co-regulated by Upf1 and PNRC2. Results provide important information that vast range of cellular SMD or NMD substrates are reqired PNRC2 for decay. Total RNA obtained from HeLa cells with downregulation of Upf1 or PNRC2 by siRNA. The up- or down-regulated transcripts were compare to control siRNA treated HeLa cell RNA extract. Significant transcripts were confirmed by replication

2012-02-28 | E-GEOD-26781 | biostudies-arrayexpress

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