Split Gal4 Biosensor for Anastasis Detection
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Solution Overview
Problem
Current cancer treatments based on apoptosis assume irreversibility, leading to cancer recurrence as dying cancer cells can recover and repopulate during treatment intervals, acquiring new mutations and contributing to drug resistance.
Innovation Solution
Development of a Split Gal4-based anastasis biosensor to detect and track the reversal of apoptosis in cancer cells, specifically labeling cells that have experienced mitochondrial outer membrane permeabilization and caspase-3 activation, allowing for the identification of cells that reverse apoptosis and potentially develop new therapeutic strategies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If cancer treatments assume apoptosis is irreversible, then treatment protocols can be simplified, but cancer cells can recover and repopulate during treatment intervals, leading to recurrence and drug resistance
Solution Approach 1:
The biosensor performs preliminary detection of apoptotic cells before they can recover and repopulate. By labeling cells that have undergone mitochondrial outer membrane permeabilization and caspase-3 activation, the system identifies vulnerable cancer cells in advance, allowing for timely intervention before recurrence occurs
Solution Approach 2:
The biosensor provides real-time feedback on the apoptotic status of cancer cells during treatment. This feedback mechanism allows clinicians to monitor treatment effectiveness and adjust protocols dynamically, preventing the recovery and repopulation of cancer cells that would otherwise lead to recurrence
2Measurement precision
If a biosensor detects both mitochondrial outer membrane permeabilization and caspase-3 activation, then anastasis events can be accurately identified, but the device complexity increases
Solution Approach 1:
The biosensor is divided into separate modular components: a mitochondrial targeting module that detects outer membrane permeabilization and a caspase-3 detection module. Each module functions independently but contributes to the overall detection capability, allowing for precise anastasis identification while maintaining manageable system complexity through functional segmentation
Data Source
AI summary
The present invention relates to the field of anastasis, i.e., the process of reversal of apoptosis. More specifically, the present invention provides methods and compositions useful for studying anastasis. The present invention also provides a biosensor comprising (a) a split transcription factor complex comprising one half of a split transcription factor linked to a transmembrane domain via an enzyme cleavable linker; (b) a split transcription factor comprising the other half of the split transcription factor linked to a MTS via an enzyme-cleavable linker; and (c) a reporter system comprising (1) a first nucleic acid encoding a site specific recombinase operably linked to the site specific sequence for the transcription factor; and (2) a second nucleic acid comprising a stop codon cassette flanked by site specific recombination sequences, wherein the split transcription factor is Gal 4 or split Q. In other embodiments, the recombinase is Cre or FLP.