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split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80
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 contr...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10055387/ https://www.ncbi.nlm.nih.gov/pubmed/36993523 http://dx.doi.org/10.1101/2023.03.24.534001 |
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author | Ewen-Campen, Ben Luan, Haojiang Xu, Jun Singh, Rohit Joshi, Neha Thakkar, Tanuj Berger, Bonnie White, Benjamin H. Perrimon, Norbert |
author_facet | Ewen-Campen, Ben Luan, Haojiang Xu, Jun Singh, Rohit Joshi, Neha Thakkar, Tanuj Berger, Bonnie White, Benjamin H. Perrimon, Norbert |
author_sort | Ewen-Campen, Ben |
collection | PubMed |
description | 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 new 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 fully 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 a new 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. |
format | Online Article Text |
id | pubmed-10055387 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Cold Spring Harbor Laboratory |
record_format | MEDLINE/PubMed |
spelling | pubmed-100553872023-03-30 split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80 Ewen-Campen, Ben Luan, Haojiang Xu, Jun Singh, Rohit Joshi, Neha Thakkar, Tanuj Berger, Bonnie White, Benjamin H. Perrimon, Norbert bioRxiv Article 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 new 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 fully 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 a new 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. Cold Spring Harbor Laboratory 2023-03-24 /pmc/articles/PMC10055387/ /pubmed/36993523 http://dx.doi.org/10.1101/2023.03.24.534001 Text en https://creativecommons.org/licenses/by/4.0/This work is licensed under a Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/) , which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use. |
spellingShingle | Article Ewen-Campen, Ben Luan, Haojiang Xu, Jun Singh, Rohit Joshi, Neha Thakkar, Tanuj Berger, Bonnie White, Benjamin H. Perrimon, Norbert split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80 |
title | split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80 |
title_full | split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80 |
title_fullStr | split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80 |
title_full_unstemmed | split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80 |
title_short | split-intein Gal4 provides intersectional genetic labeling that is fully repressible by Gal80 |
title_sort | split-intein gal4 provides intersectional genetic labeling that is fully repressible by gal80 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10055387/ https://www.ncbi.nlm.nih.gov/pubmed/36993523 http://dx.doi.org/10.1101/2023.03.24.534001 |
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