Author(s)
Thomas E. Gorochowski
Published 9 Projects
Microscopy Bioinformatics Rna Seq Synthetic Biology Systems Biology
Irina Chelysheva
Published 1 Project
Rna Seq Synthetic Biology Transcription Genetic Circuits Translation
Mette Eriksen
Published 1 Project
Rna Seq Synthetic Biology Transcription Genetic Circuits Translation
Priyanka Nair
Published 1 Project
Rna Seq Synthetic Biology Transcription Genetic Circuits Translation
Steen Pedersen
Published 1 Project
Rna Seq Synthetic Biology Transcription Genetic Circuits Translation
Zoya Ignatova
Published 1 Project
Rna Seq Synthetic Biology Transcription Genetic Circuits Translation
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Video Abstract (AI generated) (01:35) Paper Preprint Molecular Systems BiologyTranslation of mRNAs into protein is a key cellular process. Ribosome binding sites and stop codons provide signals to initiate and terminate translation, while stable secondary mRNA structures can induce translational recoding events. Fluorescent proteins, commonly used to characterize such elements, require the modification of a part's natural context and allow only a few parameters to be monitored concurrently. Here, we develop a methodology that combines ribosome profiling (Ribo-seq) with quantitative RNA sequencing (RNA-seq) to enable the high-throughput characterization of genetic parts controlling translation in absolute units. We simultaneously measure 743 translation initiation rates and 754 termination efficiencies across the Escherichia coli transcriptome, in addition to translational frameshifting induced at a stable RNA pseudoknot structure. By analyzing the transcriptional and translational response, we discover that sequestered ribosomes at the pseudoknot causes a σ32-mediated stress response, codon-specific pausing, and drop in translation initiation rates across the cell. Our work demonstrates the power of integrating global approaches to give a comprehensive and quantitative understanding of gene regulation and burden in living cells.
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Thomas E. Gorochowski. (2021, Nov 3).Absolute quantification of translational regulation and burden using combined sequencing approaches[Video]. Scitok. https://scitok.com/project/p/9ca6d547
E. Gorochowski Thomas. "Absolute quantification of translational regulation and burden using combined sequencing approaches" Scitok, uploaded by E. Gorochowski Thomas, 3 Nov, 2021, https://scitok.com/project/p9ca6d547
Thomas E. Gorochowski. "Absolute quantification of translational regulation and burden using combined sequencing approaches" Scitok. (Nov 3, 2021). https://scitok.com/project/p/9ca6d547
Thomas E. Gorochowski (Nov 3, 2021). Absolute quantification of translational regulation and burden using combined sequencing approaches Scitok. https://scitok.com/project/p/9ca6d547
Thomas E. Gorochowski. Absolute quantification of translational regulation and burden using combined sequencing approaches[video]. 2021 Nov 3. https://scitok.com/project/p/9ca6d547
@online{al2006link, title={ Absolute quantification of translational regulation and burden using combined sequencing approaches }, author={ E. Gorochowski, Thomas }, organization={Scitok}, month={ Nov }, day={ 3 }, year={ 2021 }, url = {https://scitok.com/project/p/9ca6d547}, }