The Past, Present, and Future of Genetic Manipulation in Human Fungal Pathogen Talaromyces marneffei

The fungus Talaromyces marneffei has been discovered and its pathogenicity to humans has been recognized for over sixty years. The advances in organism-wide studies and the development of genetic manipulation tools contribute greatly to our current understanding of host-pathogen interactions. Several classes of genes have been identified to be involved in stress response, morphogenesis, and virulence based on the characterization of the generated mutants. Here, we summarize the main techniques for T. marneffei genetic manipulation, including chemical mutagenesis, insertional mutagenesis, homologous recombination-mediated gene replacement, knockdown methods, and the recent popular CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats; Cas=CRISPR associated protein) technology. The advantages and disadvantages of each technique were determined from a historical perspective. We also describe potential strategies to improve the current genetics studies, such as the generation of new selection markers and genetically modified strains. The genetic approaches will continue to impact the studies of T. marneffei and can lead to the discovery of new diagnostic tools, drugs, and vaccines.