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TECHNIQUES AND RESOURCES
A targeted gene expression system using the tryptophan repressor in zebrafish shows no silencing in subsequent generations
Arminda Suli, Ali D. Guler, David W. Raible, David Kimelman
Development 2014 141: 1167-1174; doi: 10.1242/dev.100057
Arminda Suli
1 Department of Biological Structure, University of Washington, Seattle, WA 98195-5502, USA.
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Ali D. Guler
2 Department of Biochemistry, University of Washington, Seattle, WA 98195-5502, USA.
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David W. Raible
1 Department of Biological Structure, University of Washington, Seattle, WA 98195-5502, USA.
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David Kimelman
2 Department of Biochemistry, University of Washington, Seattle, WA 98195-5502, USA.
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  • For correspondence: kimelman@u.washington.edu
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Abstract

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.

  • Received June 11, 2013.
  • Accepted December 19, 2013.
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Keywords

  • Gal4/UAS
  • Gene expression system
  • Silencing
  • Tryptophan repressor
  • Zebrafish

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TECHNIQUES AND RESOURCES
A targeted gene expression system using the tryptophan repressor in zebrafish shows no silencing in subsequent generations
Arminda Suli, Ali D. Guler, David W. Raible, David Kimelman
Development 2014 141: 1167-1174; doi: 10.1242/dev.100057
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TECHNIQUES AND RESOURCES
A targeted gene expression system using the tryptophan repressor in zebrafish shows no silencing in subsequent generations
Arminda Suli, Ali D. Guler, David W. Raible, David Kimelman
Development 2014 141: 1167-1174; doi: 10.1242/dev.100057

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