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

As one of the most common airborne fungal pathogenic species, Aspergillus fumigatus infection is the increasingly fatal threat to immunocompromised patients worldwide. Setting up an efficient live-cell pathogen-labeling system will give insight into the dynamic process of the pathogen invasion in host, which offers us opportunities to explore the pathogenesis of A. fumigatus. In this chapter, we have described an efficient CRISPR-Cas9 system, which enables a precise in situ tag-insertion of an exogenous GFP tag at the predicted site with or without marker insertion in A. fumigatus. According to the detectable proportional fluorescence intensity, it is possible to in vivo track the A. fumigatus infection and to assess the fungal burden in relative organs.