Exploring a novel genomic safe-haven site in the human pathogenic mould Aspergillus fumigatus

Takanori Furukawa, Norman van Rhijn, Harry Chown, Johanna Rhodes, Narjes Alfuraiji, Rachael Fortune-Grant, Elaine Bignell, Matthew C Fisher, Michael Bromley

Research output: Contribution to journalArticlepeer-review

Abstract

Aspergillus fumigatus is the most important airborne fungal pathogen and allergen of humans causing high morbidity and mortality worldwide. The factors that govern pathogenicity of this organism are multi-factorial and are poorly understood. Molecular tools to dissect the mechanisms of pathogenicity in A. fumigatus have improved significantly over the last 20 years however many procedures have not been standardised for A. fumigatus. Here, we present a new genomic safe-haven locus at the site of an inactivated transposon, named SH-aft4, which can be used to insert DNA sequences in the genome of this fungus without impacting its phenotype. We show that we are able to effectively express a transgene construct from the SH-aft4 and that natural regulation of promoter function is conserved at this site. Furthermore, the SH-aft4 locus is highly conserved in the genome of a wide range of clinical and environmental isolates including the isolates commonly used by many laboratories CEA10, Af293 and ATCC46645, allowing a wide range of isolates to be manipulated. Our results show that the aft4 locus can serve as a site for integration of a wide range of genetic constructs to aid functional genomics studies of this important human fungal pathogen.

Original languageEnglish
Article number103702
Pages (from-to)1-10
Number of pages10
JournalFungal genetics and biology : FG & B
Volume161
Early online date18 May 2022
DOIs
Publication statusPublished - Jul 2022

Keywords

  • Aspergillus fumigatus
  • filamentous fungi
  • genome editing
  • safe-harbour site
  • safe-haven
  • transgene insertion

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