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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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Related Experiment Video

Updated: Nov 17, 2025

Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes
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Detecting circRNA in purified spliceosomal P complex.

Shasha Shi1, Xueni Li1, Rui Zhao1

  • 1Department of Biochemistry and Molecular Genetics, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, United States.

Methods (San Diego, Calif.)
|February 12, 2021
PubMed
Summary
This summary is machine-generated.

Researchers elucidated the mechanism of circular RNA (circRNA) biogenesis by identifying the spliceosomal E complex

Keywords:
Back-splicingCircRNAP complexSpliceosome

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Circular RNAs (circRNAs) are prevalent in eukaryotes and perform diverse biological functions.
  • The mechanism of circRNA formation via back-splicing remains poorly understood.
  • Canonical splicing and back-splicing share some mechanistic similarities.

Purpose of the Study:

  • To investigate the molecular mechanism underlying circRNA biogenesis.
  • To determine the role of the spliceosomal E complex in back-splicing.
  • To establish a method for studying circRNA formation and regulation.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) to determine the structure of the yeast spliceosomal E complex.
  • Purification of the yeast post-catalytic spliceosomal P complex.
  • Analysis of splicing products and intermediates from canonical and back-splicing pathways.

Main Results:

  • The spliceosomal E complex, previously studied on introns, is hypothesized to mediate exon definition for back-splicing.
  • The post-catalytic spliceosomal P complex successfully captured both canonical and back-splicing products.
  • This study provides a detailed procedure for analyzing circRNA biogenesis.

Conclusions:

  • The spliceosomal E complex is proposed to be involved in the formation of circRNAs through exon definition.
  • The described method allows for the detection of circRNA products and intermediates.
  • This research facilitates further investigation into circRNA biogenesis and regulation across species.