Jesselynn LaBelle, Adela Ramos-Martinez, Kyle Shen, Laura B Motta-Mena, Kevin H Gardner, Stefan C Materna, Stephanie Woo
{"title":"TAEL 2.0: An Improved Optogenetic Expression System for Zebrafish.","authors":"Jesselynn LaBelle, Adela Ramos-Martinez, Kyle Shen, Laura B Motta-Mena, Kevin H Gardner, Stefan C Materna, Stephanie Woo","doi":"10.1089/zeb.2020.1951","DOIUrl":null,"url":null,"abstract":"<p><p>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.</p>","PeriodicalId":23872,"journal":{"name":"Zebrafish","volume":"18 1","pages":"20-28"},"PeriodicalIF":1.4000,"publicationDate":"2021-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8020536/pdf/zeb.2020.1951.pdf","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Zebrafish","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1089/zeb.2020.1951","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2021/2/8 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"DEVELOPMENTAL BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
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.
期刊介绍:
Zebrafish is the only peer-reviewed journal dedicated to the central role of zebrafish and other aquarium species as models for the study of vertebrate development, evolution, toxicology, and human disease.
Due to its prolific reproduction and the external development of the transparent embryo, the zebrafish is a prime model for genetic and developmental studies. While genetically more distant from humans, the vertebrate zebrafish nevertheless has comparable organs and tissues, such as heart, kidney, pancreas, bones, and cartilage.
Zebrafish introduced the new section TechnoFish, which highlights these innovations for the general zebrafish community.
TechnoFish features two types of articles:
TechnoFish Previews: Important, generally useful technical advances or valuable transgenic lines
TechnoFish Methods: Brief descriptions of new methods, reagents, or transgenic lines that will be of widespread use in the zebrafish community
Zebrafish coverage includes:
Comparative genomics and evolution
Molecular/cellular mechanisms of cell growth
Genetic analysis of embryogenesis and disease
Toxicological and infectious disease models
Models for neurological disorders and aging
New methods, tools, and experimental approaches
Zebrafish also includes research with other aquarium species such as medaka, Fugu, and Xiphophorus.