Investigating the Role of TSC11, COX4, and PEP8 in Filamentation in Clinical Strains of Candida albicans
Presenter Type
UNO Undergraduate Student
Major/Field of Study
Bioinformatics
Advisor Information
Associate Professor and Chair of Biology
Location
MBSC Ballroom Poster # 504 - U
Presentation Type
Poster
Start Date
24-3-2023 10:30 AM
End Date
24-3-2023 11:45 AM
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 two clinical strains, P87 and P76067, using single guide RNA, knockout constructs, and Cas9 DNA. The deletions were confirmed by colony PCR. Our study included the genes TSC11, COX4, and PEP8, all of which are known to play an instrumental role in filamentation for the type strain SC5314. TSC11 codes for the TORC2 complex subunit, a key regulator of plasma membrane homeostasis in budding yeast cells. 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 one tested gene in one clinical strain. Confirmed knock out mutants will be assessed in filamentation assays to create more representative data for systemic infections.
Scheduling
10:45 a.m.-Noon
Investigating the Role of TSC11, COX4, and PEP8 in Filamentation in Clinical Strains of Candida albicans
MBSC Ballroom Poster # 504 - U
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 two clinical strains, P87 and P76067, using single guide RNA, knockout constructs, and Cas9 DNA. The deletions were confirmed by colony PCR. Our study included the genes TSC11, COX4, and PEP8, all of which are known to play an instrumental role in filamentation for the type strain SC5314. TSC11 codes for the TORC2 complex subunit, a key regulator of plasma membrane homeostasis in budding yeast cells. 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 one tested gene in one clinical strain. Confirmed knock out mutants will be assessed in filamentation assays to create more representative data for systemic infections.