CRISPR/Cas9 Genome Editing To Demonstrate the Contribution of Cyp51A Gly138Ser to Azole Resistance in Aspergillus fumigatus

Antimicrob Agents Chemother. 2018 Aug 27;62(9):e00894-18. doi: 10.1128/AAC.00894-18. Print 2018 Sep.

Abstract

A pan-azole-resistant Aspergillus fumigatus strain with the cyp51A mutations Gly138Ser and Asn248Lys was isolated from a patient receiving long-term voriconazole treatment. PCR fragments containing cyp51A with the mutations were introduced along with the Cas9 protein and single guide RNA into the azole-resistant/susceptible strains. Recombinant strains showed increased susceptibility via the replacement of Ser138 by glycine. Genetic recombination, which has been hampered thus far in clinical isolates, can now be achieved using CRISPR/Cas9 genome editing.

Keywords: CRISPR; Cas9; Cyp51A; antifungal resistance; azole drugs; genome editing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Antifungal Agents / therapeutic use*
  • Aspergillosis / drug therapy*
  • Aspergillus fumigatus / drug effects*
  • Aspergillus fumigatus / genetics*
  • Aspergillus fumigatus / isolation & purification
  • CRISPR-Cas Systems / genetics
  • Cytochrome P-450 Enzyme System / genetics*
  • Drug Resistance, Fungal / genetics*
  • Fungal Proteins / genetics*
  • Gene Editing / methods*
  • Humans
  • Male
  • Voriconazole / therapeutic use*

Substances

  • Antifungal Agents
  • Fungal Proteins
  • Cytochrome P-450 Enzyme System
  • cytochrome P-450 CYP51A, Aspergillus
  • Voriconazole