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Updated: Aug 12, 2025
Forward Genetic Screen Using Transgenic Calcium Reporter Aequorin to Identify Novel Targets in Calcium Signaling
Published on: August 1, 2020
Genome-wide CRISPR screen reveals genetic modifiers of Ca 2+ -mediated cell death
Abstract:
Ca 2+ is a fundamental determinant of survival in living cells. Excessive intracellular Ca 2+ causes cellular toxicity and death but the genetic pathways contributing to Ca 2+ induced cell death are incompletely understood. Here, we performed genome-wide CRISPR knock-out screening in human cells challenged with the Ca 2+ ionophore ionomycin and identified genes and pathways essential for cell death after Ca 2+ overload. We discovered 115 protective gene knockouts, 82 of which are non-essential genes and 21 of which belong to the druggable genome. Notably, members of store operated Ca 2+ entry (SOCE), very long-chain fatty acid synthesis, and SWItch/Sucrose Non-Fermentable (SWI/SNF) pathways provided marked protection against Ca 2+ toxicity. These results reveal pathways previously unknown to mediate Ca 2+ -induced cell death and provide a resource for the development of pharmacotherapies against the sequelae of Ca 2+ overload in disease.
Insights
Excessive intracellular calcium (Ca2+) causes cell death. Genome-wide screening identified protective genes and pathways, including store-operated calcium entry (SOCE), crucial for understanding and treating Ca2+ overload.
Area of Science:
- Cell Biology
- Genetics
- Biochemistry
Background:
- Calcium (Ca2+) is vital for cell survival, but excessive levels lead to toxicity and cell death.
- The genetic underpinnings of Ca2+-induced cell death remain incompletely understood, hindering therapeutic development.
Approach:
- Conducted a genome-wide CRISPR knock-out screen in human cells exposed to the Ca2+ ionophore ionomycin.
- Identified genes and pathways that confer protection against Ca2+ overload-induced cell death.
Key Points:
- Discovered 115 protective gene knockouts against Ca2+ toxicity.
- Highlighted the protective roles of store-operated calcium entry (SOCE), very long-chain fatty acid synthesis, and SWI/SNF pathways.
- Identified 21 druggable genes among the protective knockouts, offering therapeutic potential.
Conclusions:
- Revealed novel pathways involved in Ca2+-induced cell death.
- Provides a valuable resource for developing pharmacotherapies targeting diseases associated with Ca2+ overload.
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