Deus ex Candida genetics: overcoming the hurdles for the development of a molecular toolbox in the CTG clade

Emerging and Emerged Pathogenic Candida Species: Beyond the Candida albicans Paradigm

“…However, the development of genetic approaches in NAC species has been hindered by three main factors: (i) most pioneering studies during the early stages of the “pathogenic yeast genetics” field were carried out in C. albicans ; (ii) the particular codon usage of most of Candida species has precluded the direct use of S. cerevisiae or bacterial molecular tools in these NAC species [5]; (iii) most pathogenic Candida species have limited modes of sexual reproduction unlike S. cerevisiae [6].…”

“…Genetic studies of CTG clade species expanded in the 2000s and focused on the development of molecular tools, as well as transformation procedures, due to the biotechnological potential of several Candida yeasts ( C. guilliermondii , C. famata , C. tropicalis , and C. rugosa ) as well as clinical incidence ( C. dubliniensis and C. parapsilosis ) [5], [9]. Specifically, drug-resistant markers and reporter genes (encoding fluorescent protein variants, luciferase, or beta-galactosidase) were adapted by changing CTG codons to allow their functionality in this particular clade [5] (Table 1). …”

The Candida Pathogenic Species Complex

“…guilliermondii and C. lusitaniae are relatively rare causes of human infection. C. guilliermondii is of general interest to the biotechnology sector, as a producer both of riboflavin and of xylitol (Papon et al 2013b), and several molecular tools have been developed (Papon et al 2012). The species is haploid and fully sexual, which may facilitate genetic analysis.…”

Marker Recycling in Candida albicans through CRISPR-Cas9-Induced Marker Excision

“…Marker recycling has played an important role in genetic manipulation, as illustrated by the hundreds of citations to previous descriptions of marker recycling approaches (1–3). Use of these strategies is especially prominent in fungal studies, where the number of selection markers may be limited (19) and where nutritional requirements can impact diverse phenotypes. Our CRIME approach is conceptually a hybrid between the positive/negative selection strategy and the recombinase-promoted excision strategy.…”