split-intein Gal4 provides intersectional genetic labeling that is repressible by Gal80

Area of Science:

• Genetics
• Molecular Biology
• Developmental Biology

Background:

• The split-Gal4 system enables intersectional genetic labeling of specific cell types in Drosophila.
• Existing split-Gal4 systems lack temporal control as they are not repressed by Gal80, limiting their use in time-sensitive experiments.

Purpose of the Study:

• To develop a novel split-Gal4 system with temporal control and Gal80 repressibility.
• To enable precise, time-restricted genetic manipulations in Drosophila.

Main Methods:

• Development of a split-Gal4 system utilizing a self-excising split-intein.
• In vivo validation using fluorescent reporters and reversible tumor induction models.
• Integration with the GeneSwitch system for drug-inducible control.
• Application in generating cell type-specific drivers from single-cell RNA sequencing (scRNAseq) data using the TAB algorithm.
• Creation of a plasmid toolkit for driver construction via CRISPR or enhancer fragments.

Main Results:

• The novel split-intein Gal4 system demonstrates strong transgene expression comparable to existing systems.
• The system exhibits robust inducibility and repressibility by Gal80, enabling temporal control.
• Successful application in generating cell type-specific drivers predicted from scRNAseq data.
• Demonstration of reversible tumor induction in the Drosophila gut.

Conclusions:

• The split-intein Gal4 system provides a powerful tool for creating highly specific, inducible, and repressible intersectional genetic drivers in Drosophila.
• This system overcomes the temporal control limitations of previous split-Gal4 methods.
• It facilitates advanced genetic studies, including those informed by scRNAseq data analysis.