Refining GAL4-driven transgene expression in Drosophila with a GAL80 enhancer-trap
β Scribed by Maximiliano L. Suster; Laurent Seugnet; Michael Bate; Marla B. Sokolowski
- Book ID
- 102223813
- Publisher
- John Wiley and Sons
- Year
- 2004
- Tongue
- English
- Weight
- 235 KB
- Volume
- 39
- Category
- Article
- ISSN
- 1526-954X
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β¦ Synopsis
Abstract
We constructed an enhancerβtrap element, P{GAL80}, that encodes the yeast GAL80 repressor to refine expression of transgenes driven by the binary GAL4/UAS system. GAL80 blocks GAL4 activity by binding to its transcriptional activation domain. We screened GAL80 enhancerβtraps for repression of GAL4βinduced green fluorescent protein (GFP) in the intact larval nervous system. We selected one line that repressed GFP in a large set of cholinergic neurons. This line was used to refine GFP expression from a set of over 200 neurons to a subset of 20 neurons in a preselected GAL4 line. Expression of tetanus neurotoxin, a potent blocker of neurotransmitter release, in these 20 neurons reproduced an aberrant larval turning behavior previously assigned to the parental set of 200 neurons. Our results suggest that targeted GAL80 expression could become a useful means of spatially refining transgene expression in Drosophila. genesis 39:240β245, 2004. Β© 2004 WileyβLiss, Inc.
π SIMILAR VOLUMES
We have screened a collection of approximately 400 GAL4 enhancer trap lines for useful patterns of expression in the embryo, larval brain, imaginal discs, and ovary using a UAS-lacZ reporter construct. Although similar patterns of expression have previously been reported in the original P[lacZ] enha