Related Experiment Video
Updated: Jul 16, 2025
MEDUSA for Identifying Death Regulatory Genes in Chemo-genetic Profiling Data
Published on: February 7, 2025
Death-seq identifies regulators of cell death and senolytic therapies
Alex Colville1, Jie-Yu Liu2, Cristina Rodriguez-Mateo2
1Paul F. Glenn Center for the Biology of Aging and Department of Neurology and Neurological Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Genetics, Stanford University, Stanford, CA 94305, USA.
Abstract:
Selectively ablating damaged cells is an evolving therapeutic approach for age-related disease. Current methods for genome-wide screens to identify genes whose deletion might promote the death of damaged or senescent cells are generally underpowered because of the short timescales of cell death as well as the difficulty of scaling non-dividing cells. Here, we establish "Death-seq," a positive-selection CRISPR screen optimized to identify enhancers and mechanisms of cell death. Our screens identified synergistic enhancers of cell death induced by the known senolytic ABT-263. The screen also identified inducers of cell death and senescent cell clearance in models of age-related diseases by a related compound, ABT-199, which alone is not senolytic but exhibits less toxicity than ABT-263. Death-seq enables the systematic screening of cell death pathways to uncover molecular mechanisms of regulated cell death subroutines and identifies drug targets for the treatment of diverse pathological states such as senescence, cancer, and fibrosis.
Insights
A new CRISPR screen called Death-seq efficiently identifies genes that enhance cell death. This method aids in discovering senolytic drugs and targets for age-related diseases like cancer and fibrosis.
Area of Science:
- Cell biology
- Genetics
- Pharmacology
Background:
- Targeted elimination of damaged or senescent cells is a promising therapeutic strategy for age-related diseases.
- Existing genome-wide screening methods are limited by short cell death timescales and challenges in culturing non-dividing cells.
Purpose of the Study:
- To develop and validate a novel CRISPR screening platform, "Death-seq," for identifying genes that regulate cell death.
- To uncover mechanisms and enhancers of cell death, particularly in the context of senolytics and age-related pathologies.
Main Methods:
- Development of "Death-seq," a positive-selection CRISPR screen optimized for identifying cell death regulators.
- Application of Death-seq to identify synergistic enhancers of cell death induced by the senolytic compound ABT-263.
- Utilizing Death-seq to screen for inducers of cell death and senescent cell clearance with ABT-199 in models of age-related diseases.
Main Results:
- Death-seq successfully identified synergistic enhancers of ABT-263-induced cell death.
- The screen revealed ABT-199 as an inducer of cell death and senescent cell clearance, with lower toxicity than ABT-263.
- The method demonstrated efficacy in models relevant to age-related diseases.
Conclusions:
- Death-seq provides a powerful tool for systematic screening of cell death pathways.
- The study uncovered novel mechanisms and potential drug targets for treating senescence, cancer, and fibrosis.
- This approach facilitates the discovery of therapeutic strategies for diverse pathological conditions.
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