# Insertional Mutagenesis of Candida auris using Agrobacterium tumefaciens

> **NIH NIH R21** · UNIVERSITY OF TEXAS HLTH SCIENCE CENTER · 2020 · $192,500

## Abstract

Drug resistance is arguably one of the most pressing problems in infectious disease. Resistance can occur
naturally, where organisms exhbit natural low-level antibiotic susceptibility or it can occur spontaneously after
exposure to antibiotics. Fungi pose special problems to antibiotic development because there are very limited
antibiotic choices for treatment. Many promising lead compounds that are toxic to fungi unfortunately are
toxic to mamals due to the similar eukaryoctic cellular organization. Consequently antifungal development
has always been difficult for fungi. Candida auris is an emerging fungal pathogen that displays a high
frequency of natural resistance to first line antifungal treatment, but also has been shown to develop
resistance to all know antifugals. The extremely rapid spread of this fungus around the world and high
mortality rate (~30-80% depending on country) has amplified known problems in treating systemic mycosis:
the lack of antifungal choices for fungal infections. Most of what we know about antifungal drug research is
derived from studies of non-pathogenic model fungi, such as Saccharomyces cerevisiae, which are
unsuitable as pan-fungal models. To address this issue, we will develop a method that rapidly interrogates
the C. auris genome to reveal genes that are potential antifungal targets by virtue of their function being
essential to cell survival. The major objective of this study will be to develop a way to rapidly and
inexpensively identify these targets. To accomplish this goal we will develop an insertional mutagenesis
system based on the bacterial pathogen, Agrobacterium tumefaciens for C. auris. The first aim will be to
improve the existing Agrobacterium tumefaciens transformation efficiency to yield enough transformants to
produce a saturated insertional mutagenesis map. We will next develop a capture-probe based enrichment
method for recovering insertion site fragments from the predominating non-junctional genomic DNA
background. Finally, we will apply deep sequencing to these enriched fragments to identify each insertion site
and its neighboring flanking genomic DNA, and ultimately assemble a high density insertion map that will be
used to identify genes that are essential for survival, and therefore, potential antifungal targets.

## Key facts

- **NIH application ID:** 9968106
- **Project number:** 5R21AI146700-02
- **Recipient organization:** UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
- **Principal Investigator:** BRIAN WICKES
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $192,500
- **Award type:** 5
- **Project period:** 2019-07-01 → 2022-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9968106

## Citation

> US National Institutes of Health, RePORTER application 9968106, Insertional Mutagenesis of Candida auris using Agrobacterium tumefaciens (5R21AI146700-02). Retrieved via AI Analytics 2026-06-01 from https://api.ai-analytics.org/grant/nih/9968106. Licensed CC0.

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