摘要
从基础生物学研究到生物技术和基因治疗的应用,四环素调控基因表达的Tet-off和Tet-on系统被用于调节不同环境下的真核细胞基因的活性。这些系统都是基于控制细菌对四环素耐药的操控基因的活性的调控因子。Tet-off系统通过给药四环素(Tc)或四环素衍生物强力霉素(dox)使基因表达沉默,而Tet-On系统通过给药dox允许激活基因表达。由于原始的Tet系统的最初设计和构建,这些细菌衍生系统里真核细胞的功能都有了明显的提高。我们在这里回顾强力霉素调控的HIV-1变体是如何被设计以及其应用如何大大提高了Tet-On系统rtTA转录激活因子成分的的活性和对dox的敏感性。这些优化的rtTA变体的激活需要较少量的dox,这将减少副作用以及在大脑组织里让基因控制到可以达到相对较低的dox水平。
关键词: Tet-on系统,Tet-off系统、转录调控、强力霉素、基因表达、rtTA、tTA、TetR。
Current Gene Therapy
Title:Tet-On Systems For Doxycycline-inducible Gene Expression
Volume: 16 Issue: 3
Author(s): Atze T. Das, Liliane Tenenbaum, Ben Berkhout
Affiliation:
关键词: Tet-on系统,Tet-off系统、转录调控、强力霉素、基因表达、rtTA、tTA、TetR。
摘要: The tetracycline-controlled Tet-Off and Tet-On gene expression systems are used to regulate the activity of genes in eukaryotic cells in diverse settings, varying from basic biological research to biotechnology and gene therapy applications. These systems are based on regulatory elements that control the activity of the tetracycline-resistance operon in bacteria. The Tet-Off system allows silencing of gene expression by administration of tetracycline (Tc) or tetracycline-derivatives like doxycycline (dox), whereas the Tet-On system allows activation of gene expression by dox. Since the initial design and construction of the original Tet-system, these bacterium-derived systems have been significantly improved for their function in eukaryotic cells. We here review how a dox-controlled HIV-1 variant was designed and used to greatly improve the activity and dox-sensitivity of the rtTA transcriptional activator component of the Tet-On system. These optimized rtTA variants require less dox for activation, which will reduce side effects and allow gene control in tissues where a relatively low dox level can be reached, such as the brain.
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Atze T. Das, Liliane Tenenbaum, Ben Berkhout , Tet-On Systems For Doxycycline-inducible Gene Expression, Current Gene Therapy 2016; 16 (3) . https://dx.doi.org/10.2174/1566523216666160524144041
DOI https://dx.doi.org/10.2174/1566523216666160524144041 |
Print ISSN 1566-5232 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5631 |
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