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Partner independent fusion gene detection by multiplexed CRISPR-Cas9 enrichment and long read nanopore sequencing

Authors

  • C. Stangl
  • S. de Blank
  • I. Renkens
  • L. Westera
  • T. Verbeek
  • J.E. Valle-Inclan
  • R.C. González
  • A.G. Henssen
  • M.J. van Roosmalen
  • R.W. Stam
  • E.E. Voest
  • W.P. Kloosterman
  • G. van Haaften
  • G.R. Monroe

Journal

  • Nature Communications

Citation

  • Nat Commun 11 (1): 2861

Abstract

  • Fusion genes are hallmarks of various cancer types and important determinants for diagnosis, prognosis and treatment. Fusion gene partner choice and breakpoint-position promiscuity restricts diagnostic detection, even for known and recurrent configurations. Here, we develop FUDGE (FUsion Detection from Gene Enrichment) to accurately and impartially identify fusions. FUDGE couples target-selected and strand-specific CRISPR-Cas9 activity for fusion gene driver enrichment - without prior knowledge of fusion partner or breakpoint-location - to long read nanopore sequencing with the bioinformatics pipeline NanoFG. FUDGE has flexible target-loci choices and enables multiplexed enrichment for simultaneous analysis of several genes in multiple samples in one sequencing run. We observe on-average 665 fold breakpoint-site enrichment and identify nucleotide resolution fusion breakpoints within 2 days. The assay identifies cancer cell line and tumor sample fusions irrespective of partner gene or breakpoint-position. FUDGE is a rapid and versatile fusion detection assay for diagnostic pan-cancer fusion detection.


DOI

doi:10.1038/s41467-020-16641-7