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Precise and Efficient In-Frame Integration of an Exogenous GFP Tag in Aspergillus fumigatus by a CRISPR System

Chi Zhang1, Ling Lu2

  • 1Jiangsu Key laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Microbiology, College of Life Sciences, Nanjing Normal University, No.1 Wen Yuan Rd., Qi Xia District, Nanjing, 210023, China.

Methods in Molecular Biology (Clifton, N.J.)
|June 7, 2017
PubMed

Insights

Researchers developed a CRISPR-Cas9 system to label Aspergillus fumigatus with GFP. This allows for in vivo tracking of fungal infections and assessment of fungal burden in immunocompromised patients.

Area of Science:

  • Medical Mycology
  • Molecular Biology
  • Infectious Diseases

Background:

  • Aspergillus fumigatus is a common airborne fungal pathogen causing fatal infections in immunocompromised individuals.
  • Understanding the dynamic process of fungal invasion is crucial for developing effective treatments.

Purpose of the Study:

  • To establish an efficient live-cell labeling system for Aspergillus fumigatus.
  • To enable in vivo tracking and quantification of fungal infections.

Main Methods:

  • Utilized a CRISPR-Cas9 system for precise in situ tag insertion of a Green Fluorescent Protein (GFP) tag.
  • Developed a method for marker insertion alongside the GFP tag in Aspergillus fumigatus.

Main Results:

  • Successfully created a system for tagging Aspergillus fumigatus with GFP.
  • Demonstrated that detectable fluorescence intensity allows for in vivo tracking of infections.
  • Showcased the ability to assess fungal burden in host organs.

Conclusions:

  • The developed CRISPR-Cas9 system provides an efficient tool for live-cell labeling of Aspergillus fumigatus.
  • This system facilitates in vivo monitoring of fungal infections and aids in understanding pathogenesis.
  • Enables quantitative assessment of fungal burden, crucial for managing infections in immunocompromised patients.
Keywords:
Aspergillus fumigatusCRISPR-Cas9GFP

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