# Isolation of mononuclear propagules from coenocytic hyphae of the mucormycosis pathogen Rhizopus delemar

> **NIH NIH R03** · LSU HEALTH SCIENCES CENTER · 2022 · $73,500

## Abstract

1. Abstract
 Rhizopus delemar is the leading casual-agent of mucormycosis, a life-threatening fungal infection in
immunocompromised patients. Despite its clinical importance, R. delemar remains understudied due to its
limited genetic tractability and lack of available research resources. Specifically, R. delemar produces
coenocytic hyphae and multinucleated spores, resulting in a high difficulty obtaining stable gene-specific
mutant strains. We have previously employed consecutive hyphal tip microdissection, and with CRISPR-Cas9
techniques, to isolate mononuclear mutant strains. However, this isolation method is laborious and inefficient.
To circumvent this obstacle, we propose hyphal fragmentation and protoplast generation, coupled with
fluorescence-activated cell sorting (FACS), which allows isolation of mononuclear propagules to obtain mutant
clones following genetic transformation robustly. We will first optimize hyphal fragmentation and protoplast
production to enrich single nucleated cells (Aim 1a) and label R. delemar strains with fluorescent nuclear
markers through DNA transformation (Aim 1b). We will then sort and isolate mononuclear hyphal
fragments/protoplasts through FACS analysis (Aim 2a). Additionally, we will improve the DNA plasmid for
further genetic manipulation of R. delemar (Aim 2b). Our research plan's successful implementation will
provide an invaluable tool to facilitate genetic studies of R. delemar-induced mucormycosis mechanisms.

## Key facts

- **NIH application ID:** 10353429
- **Project number:** 5R03AI156254-02
- **Recipient organization:** LSU HEALTH SCIENCES CENTER
- **Principal Investigator:** PING WANG
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $73,500
- **Award type:** 5
- **Project period:** 2021-02-16 → 2024-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10353429

## Citation

> US National Institutes of Health, RePORTER application 10353429, Isolation of mononuclear propagules from coenocytic hyphae of the mucormycosis pathogen Rhizopus delemar (5R03AI156254-02). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10353429. Licensed CC0.

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