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  Native molecule sequencing by nano-ID reveals synthesis and stability of RNA isoforms

Maier, K. C., Gressel, S., Cramer, P., & Schwalb, B. (2020). Native molecule sequencing by nano-ID reveals synthesis and stability of RNA isoforms. Genome Research, 30(9), 1332-1344. doi:10.1101/gr.257857.119.

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Restricted (embargoed till 2021-03) (Max Planck Institute for Biophysical Chemistry (Karl Friedrich Bonhoeffer Institute), MBPC; )
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Maier, K. C.1, Author              
Gressel, S.1, Author              
Cramer, P.1, Author              
Schwalb, B.1, Author              
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1Department of Molecular Biology, MPI for Biophysical Chemistry, Max Planck Society, ou_1863498              

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 Abstract: Eukaryotic genes often generate a variety of RNA isoforms that can lead to functionally distinct protein variants. The synthesis and stability of RNA isoforms is poorly characterized because current methods to quantify RNA metabolism use short-read sequencing and cannot detect RNA isoforms. Here we present nanopore sequencing–based isoform dynamics (nano-ID), a method that detects newly synthesized RNA isoforms and monitors isoform metabolism. Nano-ID combines metabolic RNA labeling, long-read nanopore sequencing of native RNA molecules, and machine learning. Nano-ID derives RNA stability estimates and evaluates stability determining factors such as RNA sequence, poly(A)-tail length, secondary structure, translation efficiency, and RNA-binding proteins. Application of nano-ID to the heat shock response in human cells reveals that many RNA isoforms change their stability. Nano-ID also shows that the metabolism of individual RNA isoforms differs strongly from that estimated for the combined RNA signal at a specific gene locus. Nano-ID enables studies of RNA metabolism at the level of single RNA molecules and isoforms in different cell states and conditions.

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Language(s): eng - English
 Dates: 2020-09-042020-09
 Publication Status: Published in print
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 Rev. Type: Peer
 Identifiers: DOI: 10.1101/gr.257857.119
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Title: Genome Research
Source Genre: Journal
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Pages: - Volume / Issue: 30 (9) Sequence Number: - Start / End Page: 1332 - 1344 Identifier: -