RNA聚合酶II动态调控及其成像技术的研究进展

doi:10.13560/j.cnki.biotech.bull.1985.2020-1015

摘要/Abstract

摘要：

RNA聚合酶II（RNA polymerase II,RNAP II 或Pol II）在调节生物生长发育和响应环境变化中发挥着重要的作用。转录工厂是细胞核RNA聚合酶II集合的位点,多个基因被招募到转录工厂发生共转录,形成一个高度动态协调的过程。尽管真核生物转录过程中RNA聚合酶II的重要功能已经被广泛研究,但是植物RNAP II 在不同生理条件下的活体动态特征仍然知之甚少。活细胞单分子成像技术和分子标记技术的发展,将有助于提升人们对植物RNAP II 的活体动态特征的新认识。主要阐述了真核生物RNAP II 的结构和功能特点,并介绍了细胞核内RNAP II 的动态调控转录机制,重点总结了活细胞内RNAP II 的动态标记和成像技术的研究进展,以期为今后开展植物RNAP II 的动态调控研究提供理论依据。

关键词: 转录工厂, 动态标记技术, 单分子活体成像技术, 调控机制

Abstract:

RNA polymerase II（RNAP II or Pol II）plays a vital role in regulating the growth and development of organisms and responding to environmental changes. RNA polymerase II assembles and locates at transcription factory,where multiple genes are recruited for the formation of co-transcription,which is a highly dynamic and coordinated process. Although the critical functions of RNA polymerase II in eukaryotic gene transcription have been widely studied,the dynamic characteristics of plant RNAP II under different physiological conditions are still poorly understood. The development of single-molecule imaging and labeling techniques in living cell will contribute to improving our new understanding of the dynamic characteristics of plant RNAP II in vivo. In this review,we mainly expound the structural and functional characteristics of RNAP II,and introduce the dynamic regulation and transcription mechanism of RNAP II. Furthermore,we put the emphasis on summarizing the recent progresses in dynamic labeling and imaging techniques of RNAP II in living cells,aiming to provide theoretical basis for further studies on the dynamic regulation of RNAP II in plant.

Key words: transcription factory, dynamic labeling technology, imaging technique of single-molecule in vivo, regulatory mechanism

钱虹萍, 陈博, 林金星, 崔亚宁. RNA聚合酶II动态调控及其成像技术的研究进展[J]. 生物技术通报, 2021, 37(4): 293-302.

QIAN Hong-ping, CHEN Bo, LIN Jin-xing, CUI Ya-ning. Recent Advances on Dynamic Regulation and Imaging Techniques of RNA Polymerase II[J]. Biotechnology Bulletin, 2021, 37(4): 293-302.

图/表 3

图1 RNAP II介导的转录过程 A：转录起始;B：转录延伸;C：转录终止

Fig. 1 RNAP II mediated transcription A：Transcription initiation. B：Transcription elongation. C：Transcription termination

图2 CRISPR/Cas9和SLPs技术标记目标分子的示意图 A：CRISPR/Cas9技术标记目标分子的示意图;B：SLPs技术标记目标分子的示意图

Fig. 2 The schematic of CRISPR/Cas9 and SLPs A： The schematic of CRISPR/Cas9. B： The schematic of SLPs

表1 不同显微技术的主要优缺点

Table 1 Major merits and demerits of different microscopic technique

参数Parameter	PALM	STORM	STED	LSFM	Bayesian nanoscopy
分辨率Resolution/nm	<30	<30	20-70	亚细胞分辨率 Subcellular resolution	50 nm
成像深度Imaging depth/μm	<10	<10	<100	通常<50	通常<50
成像速度Imaging speed	慢Slow	慢-低Slow to low	中等Medium	非常快Very fast	非常快Very fast
内源性Endogenous	无No	有Yes	有Yes	有Yes	有Yes
光毒性Phototoxicity	中到高Medium to high	中等Medium	高High	低Low	低 Low
蛇币价格Equipment price	中等Medium	中等Medium	非常昂贵Very expensive	非常昂贵Very expensive	非常昂贵Very expensive

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