Enhancement of Vivid-based photo-activatable Gal4 transcription factor in mammalian cells

Cell Structure and Function, 2023 · DOI: https://doi.org/10.1247/csf.22074 · Published: December 16, 2022

Simple Explanation

The Gal4/UAS system is a widely used tool for controlling gene expression. This study focuses on improving photo-activatable (PA) versions of this system, which use light to trigger gene expression. The researchers developed a new PA-Gal4 transcription factor called eGAV, which shows improved light-induced gene expression and reduced background activity in the dark compared to existing versions. eGAV's effectiveness was demonstrated in various cell types and organisms, including cultured cells, mouse brains, and chick spinal cords.

Study Duration

Not specified

Participants

Not specified

Evidence Level

Not specified

Key Findings

1
eGAV exhibits significantly lower background transcription activity in dark conditions compared to hGAVPO, a commonly used PA-Gal4 transcription factor.
2
eGAV demonstrates improved maximum light-induced gene expression levels compared to hGAVPO.
3
eGAV enables fine regulation of downstream gene expression by adjusting blue light intensity.

Research Summary

The study aimed to develop a more reliable photo-activatable Gal4 transcription factor with improved light-induced gene expression and reduced dark leakage compared to existing systems. Through optimization of synthetic PA-Gal4 transcription factors, the researchers developed eGAV, which showed superior performance in terms of light-induced gene expression and reduced background activity. The effectiveness of eGAV was validated in various experimental systems and model organisms, demonstrating its potential as a valuable tool for spatiotemporal control of gene expression.

Practical Implications

Precise Gene Control

eGAV allows for more precise control over gene expression with reduced background noise, enhancing the reliability of light-induced gene expression studies.

Versatile Applications

eGAV can be applied to a wide range of experimental systems and model organisms, making it a versatile tool for studying dynamic gene expression changes.

Temporal Dynamics Studies

The rapid activation and deactivation kinetics of eGAV enable the study of dynamic changes in cellular gene expression, such as oscillatory patterns.

Study Limitations

1
The study focused on one-photon activation of optogenetic switches, and two-photon activation for targeted manipulation of cells in tissues or organs was not explored.
2
Linker sequences connecting Gal4 DBD, transcription AD, and VVD were not examined in detail, which could potentially improve eGAV's functions.
3
The temporal characteristics of light-induced gene expressions were sometimes different in the adapted experimental model systems.

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