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无比对的 Oxford Nanopore RNA 和 DNA 测序的 poly(A) 长度测量。

: alignment-free poly(A) length measurement for Oxford Nanopore RNA and DNA sequencing.

机构信息

Computational Biology Unit, Department of Informatics, University of Bergen, 5008 Bergen, Norway.

Sars International Centre for Marine Molecular Biology, University of Bergen, 5008 Bergen, Norway.

出版信息

RNA. 2019 Oct;25(10):1229-1241. doi: 10.1261/rna.071332.119. Epub 2019 Jul 2.

Abstract

Polyadenylation at the 3'-end is a major regulator of messenger RNA and its length is known to affect nuclear export, stability, and translation, among others. Only recently have strategies emerged that allow for genome-wide poly(A) length assessment. These methods identify genes connected to poly(A) tail measurements indirectly by short-read alignment to genetic 3'-ends. Concurrently, Oxford Nanopore Technologies (ONT) established full-length isoform-specific RNA sequencing containing the entire poly(A) tail. However, assessing poly(A) length through base-calling has so far not been possible due to the inability to resolve long homopolymeric stretches in ONT sequencing. Here we present , an R package to estimate poly(A) tail length on ONT long-read sequencing data. operates on unaligned, base-called data. It measures poly(A) tail length from both native RNA and DNA sequencing, which makes poly(A) tail studies by full-length cDNA approaches possible for the first time. We assess 's performance across different poly(A) lengths, demonstrating that is a versatile tool providing poly(A) tail estimates across a wide range of sequencing conditions.

摘要

3' 端的多聚腺苷酸化是信使 RNA 的主要调控因子,其长度已知会影响核输出、稳定性和翻译等。直到最近,才出现了允许进行全基因组多聚(A)长度评估的策略。这些方法通过短读序列比对到遗传 3' 末端,间接识别与多聚(A)尾测量相关的基因。同时,Oxford Nanopore Technologies(ONT)建立了全长异构体特异性 RNA 测序,其中包含整个多聚(A)尾。然而,由于 ONT 测序无法解析长的同源多聚体延伸,因此通过碱基调用评估多聚(A)长度到目前为止还不可能。在这里,我们提出了 ,这是一个用于在 ONT 长读测序数据上估计多聚(A)尾长度的 R 包。 无需比对,直接基于碱基调用数据运行。它可以从原始 RNA 和 DNA 测序中测量多聚(A)尾的长度,这使得首次有可能通过全长 cDNA 方法进行多聚(A)尾研究。我们评估了 在不同多聚(A)长度下的性能,证明 是一种通用工具,可在广泛的测序条件下提供多聚(A)尾估计值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1e53/6800471/6611b00e05d7/1229f01.jpg

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