An integrase toolbox to record gene-expression during plant development.

Area of Science:

• Plant biology
• Synthetic biology
• Developmental biology

Background:

• Organogenesis relies on complex, coordinated multicellular behaviors.
• Synthetic circuits are crucial for understanding in vivo signaling networks in animal development.

Purpose of the Study:

• To transfer synthetic circuit technology to plants for recording developmental processes.
• To utilize orthogonal serine integrases for site-specific DNA recombination and reporter switching in plants.

Main Methods:

• Employing orthogonal serine integrases for irreversible DNA recombination.
• Visualizing recombination via switching between fluorescent reporters.
• Integrating promoters active during lateral root initiation to amplify signals.
• Developing methods to tune integrase switching thresholds (degradation tags, nuclear localization, split-intein).

Main Results:

• Successfully transferred and implemented synthetic circuit technology in plants.
• Demonstrated permanent marking of descendant cells through integrase-mediated recombination.
• Showcased improved robustness and stability of integrase switching across generations.
• Established a toolbox for history-dependent circuit construction in plant organogenesis.

Conclusions:

• The developed integrase toolbox enables the construction of history-dependent circuits in plants.
• This technology is valuable for decoding the temporal order of gene expression during organogenesis.
• The methods enhance the reliability and stability of synthetic circuits for plant developmental studies.