# Towards a genome-wide CRISPR/Cas9 mutant library in Rhizopus delemar

> **NIH NIH R21** · LSU HEALTH SCIENCES CENTER · 2022 · $220,500

## Abstract

Rhizopus delemar is the primary cause of life-threatening mucormycosis
responsible for
approximately 70% of clinical cases, with an overall mortality rate exceeding 50%. Despite its
devastating impact on human health,
much of R. delemar’s pathobiology and molecular disease
mechanisms remains unknown. As an initial step towards constructing a high-throughput genome-wide
mutant library of R. delemar, we propose
to generate a small-scale targeted mutant library utilizing
multiplex CRISPR/Cas9 technology. In Specific Aim 1, we will synthesize a collection of single-guide
RNAs (sgRNAs) targeting 40 randomly selected genes and deliver pooled sgRNAs and Cas9 in a one-
vector construct into R. delemar. In Specific Aim 2, we will identify gene mutations using Sanger DNA
sequencing and high-throughput next-generation sequencing. We will also assess efficacy, specificity,
and off-target mutations induced by multiplex CRISPR/Cas9. The success of this exploratory research
effort will lead to the construction of whole-genome CRISPR/Cas9 mutant libraries for the systemic
examination of R. delemar pathogenesis mechanisms.

## Key facts

- **NIH application ID:** 10431481
- **Project number:** 1R21AI168867-01
- **Recipient organization:** LSU HEALTH SCIENCES CENTER
- **Principal Investigator:** PING WANG
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $220,500
- **Award type:** 1
- **Project period:** 2022-02-15 → 2024-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10431481

## Citation

> US National Institutes of Health, RePORTER application 10431481, Towards a genome-wide CRISPR/Cas9 mutant library in Rhizopus delemar (1R21AI168867-01). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10431481. Licensed CC0.

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