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

Takanori Furukawa1, Norman van Rhijn1, Harry Chown1

  • 1Manchester Fungal Infection Group, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, University of Manchester, CTF Building, 46 Grafton Street, Manchester M13 9NT, UK; Lydia Becker Institute of Immunology and Inflammation, Manchester Collaborative Centre for Inflammation Research, Division of Infection, Immunity and Respiratory Medicine, Faculty of Biology, Medicine and Health, University of Manchester, Manchester Academic Health Science Centre, Manchester, UK.

Fungal Genetics and Biology : FG & B
|May 15, 2022
PubMed

Insights

Researchers developed a new genomic safe-haven locus, SH-aft4, for Aspergillus fumigatus. This site allows stable DNA insertion for studying this important human fungal pathogen without altering its characteristics.

Area of Science:

  • Mycology
  • Molecular Biology
  • Genetics

Background:

  • Aspergillus fumigatus is a major airborne fungal pathogen causing significant human morbidity and mortality.
  • Understanding the multi-factorial pathogenicity of A. fumigatus is crucial but remains poorly understood.
  • Standardized molecular tools for genetic manipulation of A. fumigatus are limited.

Purpose of the Study:

  • To identify and characterize a novel genomic safe-haven locus in A. fumigatus for genetic manipulation.
  • To enable stable integration of DNA sequences without affecting fungal phenotype.
  • To facilitate functional genomics studies of A. fumigatus pathogenicity.

Main Methods:

  • Identification of an inactivated transposon site as a potential genomic safe-haven.
  • Insertion of transgene constructs into the identified locus (SH-aft4).
  • Assessment of transgene expression and promoter function conservation.
  • Analysis of SH-aft4 locus conservation across diverse A. fumigatus clinical and environmental isolates.

Main Results:

  • A new genomic safe-haven locus, SH-aft4, was identified and validated.
  • Transgene expression from SH-aft4 was demonstrated, with conserved natural promoter regulation.
  • The SH-aft4 locus is highly conserved in a wide range of A. fumigatus isolates, including commonly used laboratory strains.
  • The locus allows for genetic manipulation without impacting the fungal phenotype.

Conclusions:

  • The SH-aft4 locus provides a reliable site for integrating genetic constructs in A. fumigatus.
  • This tool significantly aids functional genomics research into A. fumigatus pathogenicity.
  • SH-aft4 enhances the ability to study this important human fungal pathogen across various isolates.
Keywords:
Aspergillus fumigatusFilamentous fungiGenome editingSafe-harbour siteSafe-havenTransgene insertion

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