LAFITE Reveals the Complexity of Transcript Isoforms in Subcellular Fractions

Abstract: Characterization of the subcellular distribution of RNA is essential for understanding the molecular basis of biological processes. Here, the subcellular nanopore direct RNA‐sequencing (DRS) of four lung cancer cell lines (A549, H1975, H358, and HCC4006) is performed, coupled with a computational pipeline, Low‐abundance Aware Full‐length Isoform clusTEr (LAFITE), to comprehensively analyze the full‐length cytoplasmic and nuclear transcriptome. Using additional DRS and orthogonal data sets, it is shown that LAFITE outperforms current methods for detecting full‐length transcripts, particularly for low‐abundance isoforms that are usually overlooked due to poor read coverage. Experimental validation of six novel isoforms exclusively identified by LAFITE further confirms the reliability of this pipeline. By applying LAFITE to subcellular DRS data, the complexity of the nuclear transcriptome is revealed in terms of isoform diversity, 3'‐UTR usage, m6A modification patterns, and intron retention. Overall, LAFITE provides enhanced full‐length isoform identification and enables a high‐resolution view of the RNA landscape at the isoform level.

Standort
Deutsche Nationalbibliothek Frankfurt am Main
Umfang
Online-Ressource
Sprache
Englisch

Erschienen in
LAFITE Reveals the Complexity of Transcript Isoforms in Subcellular Fractions ; day:03 ; month:12 ; year:2022 ; extent:18
Advanced science ; (03.12.2022) (gesamt 18)

Urheber
Zhang, Jizhou
Lin, Xiao
Chen, Yuelong
Li, Tsz‐Ho
Lee, Alan Chun‐Kit
Chow, Eugene Yui‐Ching
Cho, William Chi‐Shing
Chan, Ting‐Fung

DOI
10.1002/advs.202203480
URN
urn:nbn:de:101:1-2022120314183710413147
Rechteinformation
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
Letzte Aktualisierung
15.08.2025, 07:22 MESZ

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Beteiligte

  • Zhang, Jizhou
  • Lin, Xiao
  • Chen, Yuelong
  • Li, Tsz‐Ho
  • Lee, Alan Chun‐Kit
  • Chow, Eugene Yui‐Ching
  • Cho, William Chi‐Shing
  • Chan, Ting‐Fung

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