eLife (Mar 2018)
A gene-specific T2A-GAL4 library for Drosophila
- Pei-Tseng Lee,
- Jonathan Zirin,
- Oguz Kanca,
- Wen-Wen Lin,
- Karen L Schulze,
- David Li-Kroeger,
- Rong Tao,
- Colby Devereaux,
- Yanhui Hu,
- Verena Chung,
- Ying Fang,
- Yuchun He,
- Hongling Pan,
- Ming Ge,
- Zhongyuan Zuo,
- Benjamin E Housden,
- Stephanie E Mohr,
- Shinya Yamamoto,
- Robert W Levis,
- Allan C Spradling,
- Norbert Perrimon,
- Hugo J Bellen
Affiliations
- Pei-Tseng Lee
- ORCiD
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
- Jonathan Zirin
- Department of Genetics, Harvard Medical School, Boston, United States
- Oguz Kanca
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
- Wen-Wen Lin
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
- Karen L Schulze
- ORCiD
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
- David Li-Kroeger
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
- Rong Tao
- Department of Genetics, Harvard Medical School, Boston, United States
- Colby Devereaux
- Department of Genetics, Harvard Medical School, Boston, United States
- Yanhui Hu
- Department of Genetics, Harvard Medical School, Boston, United States
- Verena Chung
- Department of Genetics, Harvard Medical School, Boston, United States
- Ying Fang
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
- Yuchun He
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
- Hongling Pan
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
- Ming Ge
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
- Zhongyuan Zuo
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Jan and Dan Duncan Neurological Research Institute, Texas Children’s Hospital, Houston, United States
- Benjamin E Housden
- ORCiD
- Department of Genetics, Harvard Medical School, Boston, United States
- Stephanie E Mohr
- ORCiD
- Department of Genetics, Harvard Medical School, Boston, United States; Howard Hughes Medical Institute, Harvard Medical School, Boston, United States
- Shinya Yamamoto
- ORCiD
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Jan and Dan Duncan Neurological Research Institute, Texas Children’s Hospital, Houston, United States; Program in Developmental Biology, Baylor College of Medicine, Houston, United States; Department of Neuroscience, Baylor College of Medicine, Houston, United States
- Robert W Levis
- Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
- Allan C Spradling
- Department of Embryology, Howard Hughes Medical Institute, Carnegie Institution for Science, Baltimore, United States
- Norbert Perrimon
- ORCiD
- Department of Genetics, Harvard Medical School, Boston, United States; Howard Hughes Medical Institute, Harvard Medical School, Boston, United States
- Hugo J Bellen
- ORCiD
- Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States; Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States; Jan and Dan Duncan Neurological Research Institute, Texas Children’s Hospital, Houston, United States; Program in Developmental Biology, Baylor College of Medicine, Houston, United States; Department of Neuroscience, Baylor College of Medicine, Houston, United States
- DOI
- https://doi.org/10.7554/eLife.35574
- Journal volume & issue
-
Vol. 7
Abstract
We generated a library of ~1000 Drosophila stocks in which we inserted a construct in the intron of genes allowing expression of GAL4 under control of endogenous promoters while arresting transcription with a polyadenylation signal 3’ of the GAL4. This allows numerous applications. First, ~90% of insertions in essential genes cause a severe loss-of-function phenotype, an effective way to mutagenize genes. Interestingly, 12/14 chromosomes engineered through CRISPR do not carry second-site lethal mutations. Second, 26/36 (70%) of lethal insertions tested are rescued with a single UAS-cDNA construct. Third, loss-of-function phenotypes associated with many GAL4 insertions can be reverted by excision with UAS-flippase. Fourth, GAL4 driven UAS-GFP/RFP reports tissue and cell-type specificity of gene expression with high sensitivity. We report the expression of hundreds of genes not previously reported. Finally, inserted cassettes can be replaced with GFP or any DNA. These stocks comprise a powerful resource for assessing gene function.
Keywords
