A gene-specific T2A-GAL4 library for Drosophila

作者：Lee Pei Tseng; Zirin Jonathan; Kanca Oguz; Lin Wen Wen; Schulze Karen L; Li Kroeger David; Tao Rong; Devereaux Colby; Hu Yanhui; Chung Verena; Fang Ying; He Yuchun; Pan Hongling; Ge Ming; Zuo Zhongyuan; Housden Benjamin E; Mohr Stephanie E; Yamamoto Shinya; Levis Robert W; Spradling Allan C; Perrimon Norbert; Bellen Hugo J

来源：eLife, 2018, 7: e35574.

DOI：10.7554/eLife.35574

<jats:p>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.</jats:p>

• 出版日期2018-3-22