Tet-On Systems For Doxycycline-inducible Gene Expression.

Tet-On Systems For Doxycycline-inducible Gene Expression.
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DOI:
10.2174/1566523216666160524144041
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发表时间:
2016
影响因子:
3.6
通讯作者:
Berkhout B
Berkhout B
中科院分区:
医学4区
文献类型:
--
作者:
Das AT;Tenenbaum L;Berkhout B

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四环素控制的Tet-Off和Tet-On基因表达系统被用于在从基础生物学研究到生物技术和基因治疗应用的不同环境中调节真核细胞中基因的活性。这些系统是基于控制细菌中四环素耐药操纵子活性的调节元件。Tet-Off系统允许通过给予四环素(TC)或四环素衍生物(如强力霉素(Dox))来沉默基因表达,而Tet-on系统允许DOx激活基因表达。自从最初的Tet系统的设计和构建以来,这些细菌衍生系统在真核细胞中的功能已经得到了显著的改善。我们在这里回顾了如何设计和使用DOX控制的HIV-1变异体来极大地提高Tet-on系统的RTTA转录激活成分的活性和对Dox的敏感性。这些优化的RTTA变体需要较少的DOX才能激活,这将减少副作用,并允许在DOX水平相对较低的组织中进行基因控制,如大脑。
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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