A novel G3BP1-GFP reporter human kidney cell system enabling real-time monitoring of stress granule dynamics for in vitro kidney toxicity assessment

Abstract

Background

High salt and hyperosmolarity can cause renal cell death, which can act as a risk factor for a variety of conditions, including acute and chronic kidney disease. Therefore, a monitoring system to assess renal cytotoxicity is required.

Objectives

The study aimed to develop a system that could rapidly and accurately assess the properties of stress granules, non-membrane organelles that are regulated by liquid–liquid phase separation and form when cells are stressed, by exploiting the properties of stress granules (SGs).

Results

We established a human embryonic kidney cell line expressing endogenous Ras GTPase-activating protein-binding protein 1 (G3BP1)-green fluorescent protein (GFP) through CRISPR/Cas9 gene editing. We found that G3BP1-GFP cells formed SGs similar to native G3BP1 cells after exposure to arsenite, high salt, and osmotic stress. We also validated the human embryonic kidney cell line expressing G3BP1-GFP through real-time monitoring.

Conclusion

The G3BP1-GFP expressing human embryonic kidney cell line provides a novel method to assess kidney toxicity through real-time monitoring of SGs. This method allows for real-time monitoring of SGs in response to various renal toxicants, providing a sensitive and rapid approach for toxicity assessment.