Abstract
The Candida genus encompasses a diverse group of ascomycete fungi that have captured the attention of the scientific community, due to both their role in pathogenesis and emerging applications in biotechnology; the development of gene editing tools such as CRISPR, to analyze fungal genetics and perform functional genomic studies in these organisms, is essential to fully understand and exploit this genus, to further advance antifungal drug discovery and industrial value. However, genetic manipulation of Candida species has been met with several distinctive barriers to progress, such as unconventional codon usage in some species, as well as the absence of a complete sexual cycle in its diploid members. Despite these challenges, the last few decades have witnessed an expansion of the Candida genetic toolbox, allowing for diverse genome editing applications that range from introducing a single point mutation to generating large-scale mutant libraries for functional genomic studies. Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 technology is among the most recent of these advancements, bringing unparalleled versatility and precision to genetic manipulation of Candida species. Since its initial applications in Candida albicans, CRISPR-Cas9 platforms are rapidly evolving to permit efficient gene editing in other members of the genus. The technology has proven useful in elucidating the pathogenesis and host-pathogen interactions of medically relevant Candida species, and has led to novel insights on antifungal drug susceptibility and resistance, as well as innovative treatment strategies. CRISPR-Cas9 tools have also been exploited to uncover potential applications of Candida species in industrial contexts. This review is intended to provide a historical overview of genetic approaches used to study the Candida genus and to discuss the state of the art of CRISPR-based genetic manipulation of Candida species, highlighting its contributions to deciphering the biology of this genus, as well as providing perspectives for the future of Candida genetics.
Highlights
Among fungal species, the Candida genus has generated interest both as a threat to human health and as an asset to industrial manufacturing
This review describes the current state of the art for Clustered regularly interspaced short palindromic repeats (CRISPR)-based genetic manipulation of Candida species, with a focus on how this technology is being applied to further our understanding of the genetics of these complex fungal organisms
By using CRISPR tools to genetically engineer Candida species, pathogen-specific and essential genes involved in fitness and virulence can be identified as unique drug targets, or as adjuvants for existing antifungal therapies
Summary
The Candida genus has generated interest both as a threat to human health and as an asset to industrial manufacturing. The use of commercially available Cas9 and custom-designed RNA can be exploited as an expression-free system, and is beneficial to study Candida species where limited molecular genetic tools (plasmids, selectable markers, species-specific promoters, etc.) are available.
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