# Next-Generation ATP-Independent Luciferases, Luciferins, and Bioluminescent Calcium Indicators

> **NIH NIH R01** · UNIVERSITY OF VIRGINIA · 2024 · $461,296

## Abstract

Abstract:
In 2012, Promega introduced NanoLuc, an ATP-independent marine luciferase mutant exhibiting a high photon
production rate in the presence of a synthetic luciferin analog, furimazine. However, NanoLuc has several
unfavorable features, including low tissue penetration of its blue emission, and limited substrate solubility and
stability. Recent studies (including our efforts) have partially addressed these issues, but there are remaining
key hurdles that prevent a broader adoption of NanoLuc-derived bioluminescence systems for in vivo animal
imaging.
The overall objective of this 4-year R01 project is to develop next-generation ATP-independent luciferases,
luciferins, and bioluminescent calcium indicators to greatly facilitate imaging experiments in small animals.
Leveraging our strong preliminary results and sustained research interests and expertise in the relevant areas,
we will pursue the following three Specific Aims:
1. Develop shelf-stable and water-soluble luciferins.
2. Develop further red-shifted luciferase-luciferin pairs.
3. Engineer next-generation bioluminescent calcium indicators with enhanced physiological responsiveness.
The outcomes of this project will be bioluminescent tools to facilitate imaging studies in animal models. These
fundamental research tools will further catalyze an extensive array of biological and biomedical studies.

## Key facts

- **NIH application ID:** 10824644
- **Project number:** 1R01EB035430-01
- **Recipient organization:** UNIVERSITY OF VIRGINIA
- **Principal Investigator:** Huiwang Ai
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $461,296
- **Award type:** 1
- **Project period:** 2024-02-05 → 2028-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10824644

## Citation

> US National Institutes of Health, RePORTER application 10824644, Next-Generation ATP-Independent Luciferases, Luciferins, and Bioluminescent Calcium Indicators (1R01EB035430-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10824644. Licensed CC0.

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