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Related Concept Videos

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Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
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Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
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Identification of Alternative Splicing and Polyadenylation in RNA-seq Data
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Isoform-specific m6A deposition and coordinated splicing shape mammalian transcriptome evolution.

Gabriela Santos-Rodriguez1,2, Akanksha Srivastava3,4,5, Agin Ravindran3,4,5

  • 1Garvan Institute of Medical Research, EMBL Australia, Sydney, NSW, Australia.

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|April 21, 2026
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Summary
This summary is machine-generated.

Transcriptome evolution is shaped by alternative splicing and RNA modifications like N6-methyladenosine (m6A). Conserved isoforms and m6A sites play crucial roles in gene expression and functional diversity across mammals.

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Area of Science:

  • Genomics
  • Evolutionary Biology
  • Molecular Biology

Background:

  • Alternative isoform usage and RNA modifications are key drivers of transcriptome evolution.
  • N6-methyladenosine (m6A) is the most abundant internal mRNA modification, crucial for gene regulation.
  • Short-read sequencing limits understanding of evolutionary conservation and phenotypic impact of transcript isoforms and m6A.

Purpose of the Study:

  • To conduct a comparative evolutionary analysis of transcript isoforms and m6A modifications using direct-RNA long-read sequencing.
  • To investigate the conservation and functional significance of these elements across mammalian clades and an avian outgroup.

Main Methods:

  • Direct-RNA long-read sequencing of six tissues from multiple mammalian species and an avian outgroup.
  • Comparative analysis of transcript isoform diversity and N6-methyladenosine (m6A) modification sites.
  • Identification of conserved and species-specific isoforms and m6A modification patterns.

Main Results:

  • 71% of transcript isoforms are species-specific but contribute minimally to gene expression; 18% of conserved isoforms dominate expression.
  • 14.2% of m6A sites are conserved across mammals, enriched in 3'-UTRs and near stop codons.
  • Conserved m6A sites show isoform-specific deposition, and coordinate splicing acts as a buffering mechanism for isoform regulation.

Conclusions:

  • Post-transcriptional regulation, including alternative splicing and m6A modifications, significantly shapes phenotypic diversity and evolutionary adaptation in mammals.
  • Conserved transcript isoforms and m6A modifications are critical for maintaining gene expression and functional transcript diversity.
  • Direct-RNA long-read sequencing provides novel insights into the evolutionary dynamics of the transcriptome.