Light-controllable Transcription System by Nucleocytoplasmic Shuttling of a Truncated Phytochrome B
Light-controllable Transcription System by Nucleocytoplasmic Shuttling of a Truncated Phytochrome B
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DOI：
10.1111/php.12955
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发表时间：
2018-09-01
影响因子：
3.3
通讯作者：
Ozawa, Takeaki
中科院分区：
文献类型：
--
作者：
Noda, Natsumi;Ozawa, Takeaki
转录调控是基因治疗和生物医学研究的有效策略。与化学调控的转录方法不同，利用光遗传学工具对转录进行时空控制是一项分析单细胞的强大技术。为了使光渗透到组织中，需要使用吸收红光/远红光的光感受器，其分子质量低，适用于病毒载体的使用，并且需要构建使用光感受器的光开关作为单一表达载体。在此，我们描述了一个基于拟南芥光敏色素（Phy） B及其结合伙伴光敏色素相互作用因子（PIF） 6的光遗传学工具。我们产生了一个截断的PhyB，它允许在红光/远红光照射下与PIF6可逆关联。仅5分钟的红光照射诱导PhyB通过与PIF6的结合从细胞质转位到细胞核，导致基于Gal4 dna结合域和Gal系统上游激活序列的转录激活。利用PhyB和PIF6构建的核细胞质穿梭载体可用于组织实验中的转录调控。
Transcriptional regulation is a useful strategy for gene therapy and for biomedical research. Unlike chemically regulated transcriptional approaches, spatiotemporal control of transcription using optogenetic tools is a powerful technology for the analysis of single cells. For light to penetrate into tissues, it is desired to use photoreceptors absorbing red/far-red light with a low-molecular mass applicable for the use of virus vectors, and a photoswitch using the photoreceptor needs to be constructed as a single expression vector. Herein, we describe an optogenetic tool based on Arabidopsis thaliana phytochrome (Phy) B and its binding partner, phytochrome-interacting factor (PIF) 6. We generated a truncated PhyB, which allowed for reversible association with PIF6 by red/far-red light illumination. The red light illumination only for 5 min induced PhyB translocation from the cytoplasm into the nucleus by the association with PIF6, resulting in transcriptional activation based on Gal4 DNA-binding domain and the upstream activating sequence of Gal system. The nucleocytoplasmic shuttling vector using PhyB and PIF6 might be applicable for transcriptional regulation in tissue experiments.