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Area of Science:

  • Virology
  • Molecular Biology
  • Transcriptomics

Background:

  • The Epstein-Barr virus (EBV) transcriptome is increasingly recognized for its complexity, encompassing microRNAs, long non-coding RNAs, and polyadenylated lytic transcripts.
  • Previous studies have expanded our understanding of EBV RNA molecules, but the presence and role of viral circular RNAs remained largely unexplored.

Purpose of the Study:

  • To identify and characterize novel circular RNAs (circRNAs) encoded by Epstein-Barr virus (EBV) during both latent and lytic infection phases.
  • To investigate the expression patterns, structural features, and cellular localization of these newly discovered viral circRNAs.
  • To assess the presence of specific EBV circRNAs in clinical samples, such as EBV-positive stomach cancer.

Main Methods:

  • Utilized RNase R-sequencing to enrich for circular RNAs in cell models representing EBV latency types I, II, and III.
  • Employed sequencing and bioinformatics analyses to identify and validate EBV-encoded circular RNAs, including those formed by backsplicing events.
  • Analyzed expression profiles, nuclear/cytoplasmic distribution, and detected specific circRNAs in clinical stomach cancer specimens.

Main Results:

  • Identified over 30 unique candidate EBV circRNAs expressed during latency and reactivation.
  • Characterized circRNAs originating from latency promoters (Cp), EBNA genes, and the RPMS1 locus.
  • Detected circRNAs derived from LMP2 and BHLF1 genes during reactivation, with specific expression patterns.
  • Confirmed the presence of two RPMS1-derived EBV circRNAs in EBV-positive stomach cancer tissues.

Conclusions:

  • This study reveals a new class of viral transcripts, Epstein-Barr virus circular RNAs, expanding the known EBV transcriptome.
  • The identified circRNAs exhibit diverse expression patterns dependent on the EBV transcriptional program and cellular context.
  • The detection of EBV circRNAs in stomach cancer suggests their potential role in EBV-associated pathogenesis and warrants further functional investigation.