| Autor: |
LaBelle J; Department of Molecular Cell Biology, University of California Merced, Merced, California, USA., Ramos-Martinez A; Department of Molecular Cell Biology, University of California Merced, Merced, California, USA., Shen K; Department of Molecular Cell Biology, University of California Merced, Merced, California, USA., Motta-Mena LB; Optologix, Inc., Dallas, Texas, USA., Gardner KH; Structural Biology Initiative, CUNY Advanced Science Research Center, New York, New York, USA., Materna SC; Department of Molecular Cell Biology, University of California Merced, Merced, California, USA., Woo S; Department of Molecular Cell Biology, University of California Merced, Merced, California, USA. |
| Abstrakt: |
Inducible gene expression systems are valuable tools for studying biological processes. We previously developed an optogenetic gene expression system called TAEL that is optimized for use in zebrafish. When illuminated with blue light, TAEL transcription factors dimerize and activate gene expression downstream of the TAEL-responsive C120 promoter. By using light as the inducing agent, the TAEL/C120 system overcomes limitations of traditional inducible expression systems by enabling fine spatial and temporal regulation of gene expression. In this study, we describe ongoing efforts to improve the TAEL/C120 system. We made modifications to both the TAEL transcriptional activator and the C120 regulatory element, collectively referred to as TAEL 2.0. We demonstrate that TAEL 2.0 consistently induces higher levels of reporter gene expression and at a faster rate, but with comparable background and toxicity as the original TAEL system. With these improvements, we were able to create functional stable transgenic lines to express the TAEL 2.0 transcription factor either ubiquitously or with a tissue-specific promoter. We demonstrate that the ubiquitous line in particular can be used to induce expression at late embryonic and larval stages, addressing a major deficiency of the original TAEL system. This improved optogenetic expression system will be a broadly useful resource for the zebrafish community. |
