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Rapid Optimization of a Light-Inducible System to Control Mammalian Gene Expression
Published on: November 4, 2025
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Light-induced gene expression with photocaged IPTG for induction profiling in a high-throughput screening system
Georg Wandrey1, Claus Bier2, Dennis Binder3
1AVT-Biochemical Engineering, RWTH Aachen University, Worringerweg 1, Aachen, 52074, Germany.
Microbial Cell Factories
|April 25, 2016
Summary
This study introduces a novel light-based induction method using photocaged isopropyl β-D-1-thiogalactopyranoside (cIPTG) for optimizing heterologous protein production. This optically induced gene expression is as effective as conventional methods but offers significant advantages in high-throughput screening.
Area of Science:
- Biotechnology
- Molecular Biology
- Biochemical Engineering
Background:
- Inducible expression systems are crucial for heterologous protein production, requiring optimization of induction parameters like time and inducer concentration.
- Traditional induction profiling experiments are often laborious and time-consuming.
- A novel approach using light-induced gene expression with photocaged isopropyl β-D-1-thiogalactopyranoside (cIPTG) in a microtiter plate system is presented.
Purpose of the Study:
- To develop and evaluate a new, efficient method for induction profiling in microbial cultivations.
- To demonstrate the feasibility of using light to control gene expression via photocaged inducers.
- To optimize conditions for maximum heterologous protein production using a high-throughput screening system.
Main Methods:
- Utilized a T7-RNA polymerase-dependent E. coli expression system for flavin mononucleotide-based fluorescent reporter protein (FbFP) production.
- Employed a microtiter plate-based cultivation system (BioLector) with UV-A light-emitting diodes for controlled IPTG uncaging.
- Monitored IPTG uncaging, protein formation, and biomass growth in parallel across multiple plates.
Main Results:
- Successfully demonstrated light-induced IPTG release and subsequent protein production.
- Showed that optical induction with cIPTG yields equivalent product formation and biomass growth compared to conventional isopropyl β-D-1-thiogalactopyranoside (IPTG) induction.
- Identified optimal induction conditions, requiring minimal UV-A exposure (6-8 s) or low IPTG concentrations (60-80 µM) for maximum product formation.
Conclusions:
- Combined optical induction with online monitoring in a high-throughput screening system.
- Established light-induced gene expression with cIPTG as a cost-effective, automatable, non-invasive, and contamination-free alternative to conventional IPTG induction.
- Highlighted the potential of this method to significantly improve the efficiency of heterologous protein production optimization.
