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使用光敏核酸结合剂对转录/表达系统进行序列非依赖性和可逆光控。

Sequence-independent and reversible photocontrol of transcription/expression systems using a photosensitive nucleic acid binder.

作者信息

Estévez-Torres André, Crozatier Cécile, Diguet Antoine, Hara Tomoaki, Saito Hirohide, Yoshikawa Kenichi, Baigl Damien

机构信息

Department of Physics, Kyoto University, Kyoto 606-8502, Japan.

出版信息

Proc Natl Acad Sci U S A. 2009 Jul 28;106(30):12219-23. doi: 10.1073/pnas.0904382106. Epub 2009 Jul 14.

Abstract

To understand non-trivial biological functions, it is crucial to develop minimal synthetic models that capture their basic features. Here, we demonstrate a sequence-independent, reversible control of transcription and gene expression using a photosensitive nucleic acid binder (pNAB). By introducing a pNAB whose affinity for nucleic acids is tuned by light, in vitro RNA production, EGFP translation, and GFP expression (a set of reactions including both transcription and translation) were successfully inhibited in the dark and recovered after a short illumination at 365 nm. Our results indicate that the accessibility of the protein machinery to one or several nucleic acid binding sites can be efficiently regulated by changing the conformational/condensation state of the nucleic acid (DNA conformation or mRNA aggregation), thus regulating gene activity in an efficient, reversible, and sequence-independent manner. The possibility offered by our approach to use light to trigger various gene expression systems in a system-independent way opens interesting perspectives to study gene expression dynamics as well as to develop photocontrolled biotechnological procedures.

摘要

为了理解重要的生物学功能,开发能够捕捉其基本特征的最小合成模型至关重要。在此,我们展示了使用光敏核酸结合剂(pNAB)对转录和基因表达进行序列无关的可逆控制。通过引入一种其对核酸的亲和力可由光调节的pNAB,体外RNA产生、EGFP翻译和GFP表达(一组包括转录和翻译的反应)在黑暗中成功受到抑制,并在365nm短时间光照后恢复。我们的结果表明,通过改变核酸的构象/凝聚状态(DNA构象或mRNA聚集),可以有效调节蛋白质机器对一个或几个核酸结合位点的可及性,从而以高效、可逆和序列无关的方式调节基因活性。我们的方法以系统无关的方式利用光触发各种基因表达系统的可能性,为研究基因表达动力学以及开发光控生物技术程序开辟了有趣的前景。

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