A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells
Here, we describe a novel method based on intronic MiMIC insertions described in Nagarkar-Jaiswal et al. (2015) to perform conditional gene inactivation in Drosophila. Mosaic analysis in Drosophila cannot be easily performed in post-mitotic cells. We therefore, therefore, developed Flip-Flop, a flip...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2017-05-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/26420 |
| _version_ | 1811180671674089472 |
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| author | Sonal Nagarkar-Jaiswal Sathiya N Manivannan Zhongyuan Zuo Hugo J Bellen |
| author_facet | Sonal Nagarkar-Jaiswal Sathiya N Manivannan Zhongyuan Zuo Hugo J Bellen |
| author_sort | Sonal Nagarkar-Jaiswal |
| collection | DOAJ |
| description | Here, we describe a novel method based on intronic MiMIC insertions described in Nagarkar-Jaiswal et al. (2015) to perform conditional gene inactivation in Drosophila. Mosaic analysis in Drosophila cannot be easily performed in post-mitotic cells. We therefore, therefore, developed Flip-Flop, a flippase-dependent in vivo cassette-inversion method that marks wild-type cells with the endogenous EGFP-tagged protein, whereas mutant cells are marked with mCherry upon inversion. We document the ease and usefulness of this strategy in differential tagging of wild-type and mutant cells in mosaics. We use this approach to phenotypically characterize the loss of SNF4Aγ, encoding the γ subunit of the AMP Kinase complex. The Flip-Flop method is efficient and reliable, and permits conditional gene inactivation based on both spatial and temporal cues, in a cell cycle-, and developmental stage-independent fashion, creating a platform for systematic screens of gene function in developing and adult flies with unprecedented detail. |
| first_indexed | 2024-04-11T09:07:00Z |
| format | Article |
| id | doaj.art-4a16979eb9374174a13a109dd687cfc5 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:07:00Z |
| publishDate | 2017-05-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-4a16979eb9374174a13a109dd687cfc52022-12-22T04:32:36ZengeLife Sciences Publications LtdeLife2050-084X2017-05-01610.7554/eLife.26420A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cellsSonal Nagarkar-Jaiswal0https://orcid.org/0000-0002-2369-3714Sathiya N Manivannan1https://orcid.org/0000-0002-9470-2390Zhongyuan Zuo2Hugo J Bellen3https://orcid.org/0000-0001-5992-5989Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United StatesDepartment of Molecular and Human Genetics, Baylor College of Medicine, Houston, United StatesDepartment of Molecular and Human Genetics, Baylor College of Medicine, Houston, United StatesHoward Hughes Medical Institute, Baylor College of Medicine, Houston, United States; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Department of Neuroscience, Baylor College of Medicine, Houston, United States; Program in Developmental Biology, Baylor College of Medicine, Houston, United States; Jan and Dan Duncan Neurological Research Institute, Texas Children’s Hospital, Houston, United StatesHere, we describe a novel method based on intronic MiMIC insertions described in Nagarkar-Jaiswal et al. (2015) to perform conditional gene inactivation in Drosophila. Mosaic analysis in Drosophila cannot be easily performed in post-mitotic cells. We therefore, therefore, developed Flip-Flop, a flippase-dependent in vivo cassette-inversion method that marks wild-type cells with the endogenous EGFP-tagged protein, whereas mutant cells are marked with mCherry upon inversion. We document the ease and usefulness of this strategy in differential tagging of wild-type and mutant cells in mosaics. We use this approach to phenotypically characterize the loss of SNF4Aγ, encoding the γ subunit of the AMP Kinase complex. The Flip-Flop method is efficient and reliable, and permits conditional gene inactivation based on both spatial and temporal cues, in a cell cycle-, and developmental stage-independent fashion, creating a platform for systematic screens of gene function in developing and adult flies with unprecedented detail.https://elifesciences.org/articles/26420MiMICFLExSNF4AγeffeteTrim9post-mitotic cells |
| spellingShingle | Sonal Nagarkar-Jaiswal Sathiya N Manivannan Zhongyuan Zuo Hugo J Bellen A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells eLife MiMIC FLEx SNF4Aγ effete Trim9 post-mitotic cells |
| title | A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells |
| title_full | A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells |
| title_fullStr | A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells |
| title_full_unstemmed | A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells |
| title_short | A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells |
| title_sort | cell cycle independent conditional gene inactivation strategy for differentially tagging wild type and mutant cells |
| topic | MiMIC FLEx SNF4Aγ effete Trim9 post-mitotic cells |
| url | https://elifesciences.org/articles/26420 |
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