Poster number 54 submitted by Evan Spell
Characterization of pug1/gap2 double knockout in the human fungal pathogen C. albicans
Evan Spell (Department of Biology, Ball State University), Dr. Douglas Bernstein (Department of Biology, Ball State University)
Pseudouridine is the most common RNA modification, however its function is not well understood. Psuedouridine is broken down in many organisms by pseudouridine glycosidases, yet in humans breakdown is absent. Pseudouridine glucosidases have not been characterized in any eukaryotes. To characterize the role of psuedouridine glycosidase (PUG1) it was knocked out in Candia albicans, a eukaryotic model organism and human fungal pathogen. When the gene coding for psuedouridine glycosidase was deleted, a number of genes were up and down regulated. One of these genes GAP2, an amino acid permease, was upregulated in the absence of PUG1. The purpose of this study is to characterize the role of these genes that interact with PUG1, by deleting them, and characterizing associated phenotypes. We first generated CRISPR-Cas9 mediated mutagenesis vectors that targeted GAP2. These plasmids were then transformed into the C. albicans pug1∆ along with a repair template that introduced a stop codon in the GAP2 open reading frame. This process results in a pug1/gap2 double knockout. We are now in the process of creating additional knockout strains, including a GAP2 single deletion. These strains will then be used in comparative experiments that will characterize viability and virulence of the pug1/gap2 double knockout.
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Keywords: Knockout, C albicans