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A high-throughput, homogeneous microplate assay for agents that kill mammalian tissue culture cells
Michael Pierce1, Chunwei Wang, Matt Rebentisch
1Deltagen Proteomics, Salt Lake City, UT 84108, USA.
Abstract:
Screens for cytostasis/cytoxicity have considerable value for the discovery of therapeutic agents and the investigation of the biology of apoptosis. For instance, genetic screens for proteins, protein fragments, peptides, RNAs, or chemicals that kill tissue culture cells may aid in identifying new cancer therapeutic targets. A microplate assay for cell death is needed to achieve throughputs sufficient to sift through thousands of agents from expression or chemical libraries. The authors describe a homogeneous assay for cell death in tissue culture cells compatible with 96- or 384-well plates. In combination with a previously described system for retroviral packaging and transduction, nearly 6000 expression library clones could be screened per week in a 96-well plate format. The screening system may also prove useful for chemical screens.
Insights
This study presents a high-throughput microplate assay for detecting cell death, crucial for discovering new cancer therapeutics. The assay enables rapid screening of thousands of agents, accelerating the identification of potential drug candidates.
Area of Science:
- Cell biology
- Molecular biology
- Drug discovery
Background:
- Cytostasis/cytotoxicity screens are vital for identifying therapeutic agents and understanding apoptosis.
- Genetic screens can uncover novel cancer therapeutic targets by identifying cell-killing agents.
- High-throughput screening methods are needed to analyze large libraries of potential therapeutic compounds.
Purpose of the Study:
- To develop a homogeneous microplate assay for detecting cell death in tissue culture.
- To enable high-throughput screening of expression and chemical libraries for therapeutic agents.
- To facilitate the identification of new cancer therapeutic targets.
Main Methods:
- A homogeneous microplate assay for cell death was developed.
- The assay is compatible with 96- and 384-well plate formats.
- The assay was integrated with a retroviral packaging and transduction system for genetic screens.
Main Results:
- The assay allows for the detection of cell death in tissue culture cells.
- Nearly 6000 expression library clones can be screened weekly using a 96-well plate format.
- The screening system demonstrates utility for both genetic and chemical screens.
Conclusions:
- The developed homogeneous assay significantly enhances the throughput of cell death screening.
- This method accelerates the discovery of therapeutic agents and cancer targets.
- The assay system is versatile and applicable to both genetic and chemical screening approaches.