Investigating the role of PHR1, COX4, and PEP8 in filamentation for clinical strains of Candida albicans
Abstract
Candia albicans is the most common fungal organism within the microbiome in humans, typically existing in the mouth, throat, and gut. However, in the right environment, C. albicans can overgrow and lead to candidiasis presenting as oral thrush or vaginal infections. Upon spreading to the blood, heart, or brain, systemic infections can have a mortality rate of approximately 25%. These infections are difficult to treat, and there are limited drugs available to treat systemic fungal infections. The ability of C. albicans to transition between rounded yeast-like cells and elongated filamentous cells is linked to pathogenesis. In this study, we compared mutated clinical strains to the type strain, SC5314, in filamentation assays to assess the diversity in the presentation of clinical C. albicans infections. These clinical strains have known phenotypic and genotypic differences from SC5314. First, novel knock out mutants of our genes of interest were generated across one clinical strain, P87, using single guide RNA, knockout constructs, and Cas9 DNA. The deletions were confirmed by colony PCR. Our study included the genes PHR1, COX4, and PEP8, all of which are known to play an instrumental role in filamentation for the type strain SC5314. PHR1 is heavily regulated by extracellular pH and plays a crucial role in cell wall assembly. COX4 functions as a large transmembrane protein and is influential in the electron transport chain and may have a role in hyphal formation. PEP8 acts as the retromer subunit, which is vital to the endosomal protein sorting machinery. All these genes contain a potential key in the infection perseverance of C. albicans, and we plan to test if these genes have the same influence on filamentation in clinical strains as SC5314. So far, we have seen growth on selective media for six tested genes in the clinical strain and three in SC5314. Confirmed knock out mutants will be assessed in filamentation assays to create more representative data for systemic infections.
Investigating the role of PHR1, COX4, and PEP8 in filamentation for clinical strains of Candida albicans
CEC RM #201/205/209
Candia albicans is the most common fungal organism within the microbiome in humans, typically existing in the mouth, throat, and gut. However, in the right environment, C. albicans can overgrow and lead to candidiasis presenting as oral thrush or vaginal infections. Upon spreading to the blood, heart, or brain, systemic infections can have a mortality rate of approximately 25%. These infections are difficult to treat, and there are limited drugs available to treat systemic fungal infections. The ability of C. albicans to transition between rounded yeast-like cells and elongated filamentous cells is linked to pathogenesis. In this study, we compared mutated clinical strains to the type strain, SC5314, in filamentation assays to assess the diversity in the presentation of clinical C. albicans infections. These clinical strains have known phenotypic and genotypic differences from SC5314. First, novel knock out mutants of our genes of interest were generated across one clinical strain, P87, using single guide RNA, knockout constructs, and Cas9 DNA. The deletions were confirmed by colony PCR. Our study included the genes PHR1, COX4, and PEP8, all of which are known to play an instrumental role in filamentation for the type strain SC5314. PHR1 is heavily regulated by extracellular pH and plays a crucial role in cell wall assembly. COX4 functions as a large transmembrane protein and is influential in the electron transport chain and may have a role in hyphal formation. PEP8 acts as the retromer subunit, which is vital to the endosomal protein sorting machinery. All these genes contain a potential key in the infection perseverance of C. albicans, and we plan to test if these genes have the same influence on filamentation in clinical strains as SC5314. So far, we have seen growth on selective media for six tested genes in the clinical strain and three in SC5314. Confirmed knock out mutants will be assessed in filamentation assays to create more representative data for systemic infections.