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Related Concept Videos

Ribosome Profiling02:24

Ribosome Profiling

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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
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Related Experiment Video

Updated: Oct 23, 2025

Identification of Alternative Splicing and Polyadenylation in RNA-seq Data
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Isoform Age - Splice Isoform Profiling Using Long-Read Technologies.

Ricardo De Paoli-Iseppi1, Josie Gleeson1, Michael B Clark1

  • 1Centre for Stem Cell Systems, Department of Anatomy and Physiology, The University of Melbourne, Parkville, VIC, Australia.

Frontiers in Molecular Biosciences
|August 19, 2021
PubMed
Summary
This summary is machine-generated.

Long-read sequencing (LRS) advances the study of alternative splicing (AS) by enabling full transcript coverage. This technology helps resolve complex RNA isoforms, offering new insights into disease mechanisms and potential therapeutic targets.

Keywords:
Oxford Nanopore Technologies nanopore sequencingPacBioalternative splicingisoformlong-read sequencingsingle cell sequencingspatial transcriptomicstargeted RNA sequencing

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Area of Science:

  • Molecular Biology
  • Genomics
  • Transcriptomics

Background:

  • Alternative splicing (AS) generates diverse RNA isoforms from single genes, increasing proteomic and transcriptomic complexity.
  • Dysregulation of AS is implicated in diseases like cancer and complex genetic disorders.
  • Short-read sequencing has limitations in resolving complex isoforms.

Purpose of the Study:

  • To review how long-read sequencing (LRS) technologies address the limitations of short-read sequencing in alternative splicing analysis.
  • To highlight the integration of LRS with other advanced techniques for detailed isoform profiling.
  • To discuss the impact of AS on protein function and its potential as a therapeutic target.

Main Methods:

  • Review of current literature on long-read sequencing (LRS) applications in alternative splicing (AS).
  • Discussion of integrated approaches combining LRS with single-cell, targeted, and spatial transcriptomics.
  • Examination of LRS coupled with genotype data to assess genetic variation's impact on isoform expression.

Main Results:

  • LRS provides full transcript coverage, enabling accurate resolution of complex RNA isoforms.
  • Integrated LRS approaches offer unprecedented detail in profiling alternative splicing at the isoform level.
  • LRS facilitates the determination of genetic variation's influence on isoform expression.

Conclusions:

  • LRS technologies are crucial for advancing the understanding of alternative splicing complexity and its role in biological pathways and diseases.
  • The integration of LRS with other sequencing modalities provides powerful tools for isoform-level analysis.
  • Understanding AS through LRS can uncover novel therapeutic targets for various diseases.