期刊
ELIFE
卷 7, 期 -, 页码 -出版社
ELIFE SCIENCES PUBLICATIONS LTD
DOI: 10.7554/eLife.35574
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资金
- National Institutes of Health [R01GM067858, U54NS093793, R24OD02205]
- National Institute of General Medical Sciences [GM067761, GM084947]
- Howard Hughes Medical Institute
- Dana-Farber/Harvard Cancer Center [5 P30 CA06516]
- Huffington Foundation
- Alzheimer's Association [NIRH-15-364099]
- Simons Foundation [368479]
- Naman Family Fund for Basic Research
- Caroline Wiess Law Fund
- Eunice Kennedy Shriver National Institute of Child Health and Human Development [U54HD083092]
- Robert A. and Renee E. Belfer Family Foundation
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, similar to 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.
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