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• W4379467121 abstract "The split-Gal4 system allows for intersectional genetic labeling of highly specific cell types and tissues in Drosophila . However, the existing split-Gal4 system, unlike the standard Gal4 system, cannot be repressed by Gal80, and therefore cannot be controlled temporally. This lack of temporal control precludes split-Gal4 experiments in which a genetic manipulation must be restricted to specific timepoints. Here, we describe a split-Gal4 system based on a self-excising split-intein, which drives transgene expression as strongly as the current split-Gal4 system and Gal4 reagents, yet which is repressible by Gal80. We demonstrate the potent inducibility of “split-intein Gal4” in vivo using both fluorescent reporters and via reversible tumor induction in the gut. Further, we show that our split-intein Gal4 can be extended to the drug-inducible GeneSwitch system, providing an independent method for intersectional labeling with inducible control. We also show that the split-intein Gal4 system can be used to generate highly cell type–specific genetic drivers based on in silico predictions generated by single-cell RNAseq (scRNAseq) datasets, and we describe an algorithm (“Two Against Background” or TAB) to predict cluster-specific gene pairs across multiple tissue-specific scRNA datasets. We provide a plasmid toolkit to efficiently create split-intein Gal4 drivers based on either CRISPR knock-ins to target genes or using enhancer fragments. Altogether, the split-intein Gal4 system allows for the creation of highly specific intersectional genetic drivers that are inducible/repressible." @default.
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• W4379467121 date "2023-06-05" @default.
• W4379467121 modified "2023-10-16" @default.
• W4379467121 title "split-intein Gal4 provides intersectional genetic labeling that is repressible by Gal80" @default.
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• W4379467121 doi "https://doi.org/10.1073/pnas.2304730120" @default.
• W4379467121 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/37276389" @default.
• W4379467121 hasPublicationYear "2023" @default.
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