A targeted gene expression system using the tryptophan repressor in zebrafish shows no silencing in subsequent generations.
- Authors
- Suli, A., Guler, A.D., Raible, D.W., Kimelman, D.
- ID
- ZDB-PUB-170525-2
- Date
- 2014
- Source
- Development (Cambridge, England) 141(5): 1167-74 (Journal)
- Registered Authors
- Kimelman, David, Raible, David, Suli, Arminda
- Keywords
- Gal4/UAS, Gene expression system, Silencing, Tryptophan repressor, Zebrafish
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Bacterial Proteins/genetics*
- Gene Silencing/physiology*
- Repressor Proteins/genetics*
- Zebrafish
- PubMed
- 24550120 Full text @ Development
The ability to visualize and manipulate cell fate and gene expression in specific cell populations has made gene expression systems valuable tools in developmental biology studies. Here, we describe a new system that uses the E. coli tryptophan repressor and its upstream activation sequence (TrpR/tUAS) to drive gene expression in stable zebrafish transgenic lines and in mammalian cells. We show that TrpR/tUAS transgenes are not silenced in subsequent generations of zebrafish, which is a major improvement over some of the existing systems, such as Gal4/gUAS and the Q-system. TrpR transcriptional activity can be tuned by mutations in its DNA-binding domain, or silenced by Gal80 when fused to the Gal4 activation domain. In cases in which more than one cell population needs to be manipulated, TrpR/tUAS can be used in combination with other, existing systems.