A Simple and Universal System for Gene Manipulation in Aspergillus fumigatus: <i>In Vitro</i> -Assembled Cas9-Guide RNA Ribonucleoproteins Coupled with Microhomology Repair Templates

DOI Web Site 6 Citations
  • Qusai Al Abdallah
    Department of Clinical Pharmacy and Translational Science, University of Tennessee Health Science Center, Memphis, Tennessee, USA
  • Wenbo Ge
    Department of Clinical Pharmacy and Translational Science, University of Tennessee Health Science Center, Memphis, Tennessee, USA
  • Jarrod R. Fortwendel
    Department of Clinical Pharmacy and Translational Science, University of Tennessee Health Science Center, Memphis, Tennessee, USA
  • Aaron P. Mitchell
    editor

Description

<jats:p> Tackling the multifactorial nature of virulence and antifungal drug resistance in <jats:named-content content-type="genus-species">A. fumigatus</jats:named-content> requires the mechanistic interrogation of a multitude of genes, sometimes across multiple genetic backgrounds. Classical fungal gene replacement systems can be laborious and time-consuming and, in wild-type isolates, are impeded by low rates of homologous recombination. Our simple and universal CRISPR-Cas9 system for gene manipulation generates efficient gene targeting across different genetic backgrounds of <jats:named-content content-type="genus-species">A. fumigatus</jats:named-content> . We anticipate that our system will simplify genome editing in <jats:named-content content-type="genus-species">A. fumigatus</jats:named-content> , allowing for the generation of single- and multigene knockout libraries. In addition, our system will facilitate the delineation of virulence factors and antifungal drug resistance genes in different genetic backgrounds of <jats:named-content content-type="genus-species">A. fumigatus</jats:named-content> . </jats:p>

Journal

  • mSphere

    mSphere 2 (6), e00446-, 2017-12-27

    American Society for Microbiology

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