Toward construction of a self-sustained clock-like expression system based on the mammalian circadian clock
Toward construction of a self-sustained clock-like expression system based on the mammalian circadian clock
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DOI：
10.1002/bit.20143
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发表时间：
2004-07-20
影响因子：
3.8
通讯作者：
Fussenegger, M
中科院分区：
文献类型：
--
作者：
Chilov, D;Fussenegger, M
尽管最近在昼夜生物学方面取得了进展，但详细了解生物起搏器系统是如何组装、维护和调节的仍然是一个重大的挑战。我们已经组装并鉴定了第一代、可调节的、自我维持的时钟样表达系统，该系统基于哺乳动物生物钟的关键组件。钟状振荡器的分子结构被简化为所有已知的昼夜节律起搏器共有的正负元素的核心集合。复杂的四环素反应多顺反子表达与最先进的慢病毒转导工具相结合，能够在人类细胞系中自动调节报告基因的表达。我们研究了一种人工振荡器的转基因表达动力学，并表明其表达谱可以通过血清休克和给药调节四环素抗生素来调节。设计一种通用的哺乳动物钟状表达系统将为研究昼夜生物学提供新的机会，并可能为有节奏地表达所需的转基因提供独特的工具，促进生物制药制造、基因治疗和组织工程的进步。(C)2004年威利期刊公司。
Despite recent advances in circadian biology, detailed understanding of how a biological pacemaker system is assembled, maintained, and regulated continues to be a significant challenge. We have assembled and characterized a first-generation, regulatable, self-sustained clock-like expression system based on key components of the mammalian circadian clock. The molecular setup of the clock-like oscillator was reduced to the core set of positive and negative elements common to all known circadian pacemakers. Sophisticated tetracycline-responsive multi-cistronic expression integrated with forefront lentiviral transduction tools enabled autoregulated reporter transgene expression in a human cell line. We characterized transgene expression kinetics of an artificial oscillator and showed that its expression profiles could be modulated by a serum shock and administration of regulating tetracycline antibiotics. Design of a generic mammalian clock-like expression system will offer novel opportunities to study circadian biology and may provide a unique tool for rhythmic expression of desired transgenes fostering advances in biopharmaceutical manufacturing, gene therapy, and tissue engineering. (C) 2004 Wiley Periodicals, Inc.